Manufacturing within a single-use container

A manufacturing assembly has at least a sterilizable chamber containing at least one of a three-dimensional printing device (additive manufacturing), a Computer Numerical Controlled (CNC) finishing head (subtractive manufacturing), a vacuum-forming unit, an injection-molding unit, a laser-cutting unit, a ultrasonic-welding unit, a robotic arman analysis device, a sampling device or a combination thereof. A plurality of individual sterilizable chambers may be aseptically connected into a network of sterilizable chambers that provides additional functionality for the manufacturing assembly. A sterilizable printer assembly may include at least one printing head, a printing platform, and a driving mechanism adapted to perform a movement of the at least one printing head relative to the printing platform along three degrees of freedom; a printer housing enclosing the printer assembly in a sterile manner, at least one aseptic connector fluidly connected to a corresponding one of the at least one printing head.

BACKGROUND

1. Field of the Invention

The invention relates to manufacturing methods for use inside of a sterile/sterilizable single-use three-dimensional container or chamber, manipulation of the manufactured products, and coating, growing, or integrating biologically active materials with those manufactured products.

2. Description of the Related Art

The forming of three-dimensional objects or the coating of three-dimensional objects under sterile conditions, particularly a coating with biochemical materials, requires high effort to uphold the sterile conditions and to assemble the required apparatuses for forming or coating under sterile conditions.

Thus, it is a problem to provide a manufacturing device, a manufacturing system and manufacturing methods that are capable to form, cut, print, or coat a material under sterile conditions in a reliable and easy manner.

SUMMARY OF THE INVENTION

The manufacturing operations performed in the systems and/or devices described in the following may be performed relative to a sterile/sterilizable chamber, in that the operations may take place within the sterilized chamber or be carried out in relation to the dimensions or walls of the chamber. One aspect of the invention relates to a manufacturing device that is a three-dimensional printing device comprising a sterilizable printer assembly including at least one printing head, a printing platform, and a driving mechanism adapted to achieve a relative displacement between the at least one printing head and the printing platform along two or three degrees of freedom. A printer housing encloses the printer assembly in a sterile manner, and at least one aseptic connector is fluidly connected to a corresponding one of the at least one printing head.

The printer assembly is intended for single-use applications, and, thus, is disposable after use. Preferably, the three-dimensional printing device is disposable with the inclusion of the printer housing after a single use. The printer housing can comprise a rigid or flexible wall, and is preferably made of a sterilizable material. An external structural assembly, such as a stainless steel support container, may be utilized to provide rigid support to a flexible-walled printer housing. An example of a printer housing may be a sterile or sterilizable chamber.

The printer assembly is sterile or sterilizable, e.g. by gamma-irradiation, autoclaving, or chemical sterilant such as ethylene oxide or vaporized hydrogen peroxide.

The printer assembly may comprise a single printing head or a plurality of printing heads. Each of the printing heads can utilize a specific technique of additive manufacturing such as extrusion, fused deposition modeling, heated extrusion, spray deposition, granular material binding, photopolymerization, etc. The printing heads are adapted for the deposition of the materials onto a printing platform or printing tray or any object located thereon. A three-dimensional computer-aided design (CAD) file can be utilized to serve as the instructions for building the three-dimensional printed structure. By depositing the appropriate printing material, three-dimensional objects may be created or formed. Other appropriate printing material may be used for coating two-dimensional or three-dimensional objects. The printer assembly may be utilized for the dispensing of nutrient media or the metered dosing of at least one drug product.

In order to print the three-dimensional object, it can be sufficient to allow a relative displacement between the at least one printing head and the printing platform along two degrees of freedom, wherein the third dimension of the object is created during by depositing printing material. A displacement of the printing platform and the at least one printing head along three degrees of freedom is an option. In order to obtain the relative displacement the at least one printing head or the printing platform can be moved. Particularly, the driving mechanism is adapted to perform a movement of the at least one printing head relative to the printing platform along at least two or three degrees of freedom. Alternatively, the driving mechanism is adapted to perform a movement of the printing platform relative to the at least one printing head along two or three degrees of freedom.

A finishing head can be utilized to subtractively remove material from the three-dimensional object after the additive manufacturing steps have been completed. Debris generated from the subtractive process can be flushed into a debris tray with a fluid media and filtered out during a recirculation of the fluid within the debris tray.

Optionally, the aseptic connector of the three-dimensional printer comprises a single-use aseptic connector, where a sterile connection and the sterile fluid transfer can occur between at least two connected complementary aseptic connectors. The aseptic connector can additionally comprise a thermoplastic tubing, which can be heat-connected in a sterile manner to a complementary aseptic connector. The heat-connection can be performed by at least one of welding, ultrasonic welding, partial melting and gluing with a thermoplastic glue.

Optionally, the internal volume of the printer housing is fluidly connected with the exterior by means of a venting filter. The venting filter may be a sterilizing grade vent filter in order to keep the interior of the printer housing in a sterile condition. The interior of the printer housing enclosing the printer assembly can be filled with a homogenous or heterogeneous gas mixture such as compressed air, nitrogen, carbon dioxide, or other mixtures and can be vented to the exterior utilizing a venting filter, preferably a sterilizing-grade vent filter.

Optionally, the at least one printing head is in a fixed position and the printing platform can be moved along two or three degrees of freedom. A degree of freedom can be a movement along a linear axis or a rotation around a rotational axis. Thus, the three degrees of freedom may comprise three rotational axes or may comprise two rotational axes and one linear axis or may comprise one rotational axis and two linear axes or may comprise three linear axes. Preferably, the degrees of freedom are all linear axes, which are orthogonal to each other, and thus, the axes are defining a cartesian coordinate system.

Optionally, the at least one printing head is in a fixed position and the printing platform can be moved along multiple degrees of freedom, such as more than three degrees of freedom or more than six degrees of freedom. Multiple degrees of freedom for the printing platform and/or the printing head can be achieved by linkage mechanisms and/or joints, which enable articulation along a plurality of angles by combining movements along a linear axis and/or rotations around a rotational axis of multiple rigid bodies connected together. Thus, the degrees of freedom may be increased to allow manufacturing by printing at a plurality of angles within a three-axis coordinate system.

Optionally, the at least one printing head is in a fixed position and contains a plurality of articulating joints which may position the dispensing printing head along multiple degrees of freedom relative to the printing platform.

Optionally, the at least one printing head of the three-dimensional printer is in a fixed position and the printing platform can be moved along multiple degrees of freedom. The printing platform may be moved along the axis framework utilizing a pneumatic actuator, a hydraulic actuator, an electric actuator, or a magnetic actuator.

Optionally, the pneumatic/hydraulic actuator comprises at least one bag or bladder, which is extendible in at least one direction by providing a fluid pressure to the inside of the at least one bag or bladder. In order to obtain a precise movement and positioning of the printing platform by means of the pneumatic/hydraulic actuator, the actuator may comprise a coarse bag and a fine bag, wherein the coarse bag is more extendible in at least one direction than the fine bag, when filled with an identical volume of a fluid or when the same internal fluid pressure is applied to the bags. Additionally, the printing platform can contain a plurality of coarse and fine bags, which can be utilized to automatically level and calibrate the distance between the printing platform and the printing head in use.

Optionally, the pneumatic/hydraulic actuator may comprise at least one of linear actuators, rolling diaphragm actuators, sterilizable linear actuators, sterilizable syringes, compressible cylinder, hydraulic rotary actuator, and hydraulic rotary vane actuators that are extendible in at least one direction by providing a fluid pressure to the inside of the actuating unit. In order to obtain a precise movement and positioning of the printing platform by means of the pneumatic/hydraulic actuator, the actuator may comprise a coarse actuator and a fine actuator, wherein the coarse actuator is more extendible in at least one direction than the fine actuator, when filled with an identical volume of a fluid or when the same internal fluid pressure is applied to the actuators. Additionally, the printing platform can contain a plurality of coarse and fine actuators, which can be utilized to automatically level and calibrate the distance between the printing platform and the printing head in use. To reduce the friction between components, the actuators may utilize rolling diaphragms to reduce the loss of force transferred via friction and to respond to changes in force with higher precision and more immediate feedback.

As an option, the pneumatic/hydraulic actuator may push and/or pull at least one printing tray across the printing platform along a plurality of tracks. The plurality of tracks may serve as guides for the positioning of the at least one printing tray platform relative to the at least one printing head.

As an option, the pneumatic/hydraulic actuator may comprise at least one fluidactuated motor that moves along a track or a threaded screw for positioning the printing platform along a corresponding axis. The fluid actuating the motor may comprise any gas or gas-liquid-mixture. Particularly, the fluid can comprise air, nitrogen, or any inert gas.

As a further option, the three-dimensional printer can comprise a magnetic actuator comprising at least one magnetic drive mechanism, which is rotatable or linearly movable by a complementary external magnetic drive mechanism.

As a further option, the three-dimensional printer may comprise a plurality of actuators formed into a robotic arm for positioning the printing platform at a plurality of positions and a plurality of angles relative to the at least one printing head.

As a further option, the three-dimensional printer may comprise a platform jack, which may convert rotational force into a linear motion. The platform jack may be utilized to control the positioning of the printing platform or printing tray. The platform jack screw may be rotatable by a complementary external magnetic drive mechanism. An internal extension assembly may be utilized to maintain a connection to transfer the force from the external assembly to the platform jack screw even as the screw position is changed by the movement of the assembly during extension and/or retraction.

As a further option, the three-dimensional printer may comprise a plurality of soft robotic actuators, where the soft material is actuated when pressurized with hydraulic and/or pneumatic pressure. The soft robotic actuators may be formed to move the at least one printing tray along the printing platform and/or move the at least one printing head into position above the at least one printing tray.

As a further option, the three-dimensional printer may comprise at least one untethered multiaxis printing device, where the device is self-contained and internal to the sterile chamber. The untethered multiaxis printing device positions itself on a static and/or fixed printing tray on the printing platform. The untethered multiaxis printing device may contain a plurality of fluid reservoirs and an internal drive mechanism to power the positional movement of the device above the printing tray.

Optionally, the three-dimensional printer comprises a position tracking system, also called precision tracking system, which is capable to determine the position of the printing platform relative to the at least one printing head along each of the three degrees of freedom. Thus, one or more of the coordinates of the printing platform within the three axis framework can be measured precisely by means of the tracking system. In order to obtain a maximum accuracy of positioning the printing platform along one or more of the framework axes a plurality of (magnetic, hydraulic, or pneumatic) actuators can be utilized along the same axis of freedom. Particularly, two or more actuators may actuate a corresponding track or threaded screw in order to provide coarse to fine resolution for movement of the printing platform. A position controller may be provided to control and auto-correct the movement of the actuators to position the printing platform to the exact coordinates required for the deposition of material by the at least one printing head.

The tracking system can include a laser tracking system, comprising an external laser source arranged exterior to the three-dimensional printer assembly, a mirror or reflective material on at least one portion of the printing platform, a laser detecting device, for detecting the time and angle of the reflected laser emission, and a computing system to calculate and report the coordinates of the printing platform relative to the at least one printing head.

Alternatively, the tracking device can comprise a plurality of cameras exterior to the three-dimensional printer assembly, a visual target material on at least one portion of the printing platform, and a computing system to calculate and report the coordinates of the printing platform relative to the at least one printing head. The cameras may be macro cameras with high resolution, such as HDTV cameras or generally cameras having a horizontal resolution of 1000 pixels or more.

Optionally, the three-dimensional printer comprises a transfer hatch adapted for removing the printing platform containing the three-dimensional printed object. By means of the transfer hatch objects may be transferred from or into the printer housing in a sterile manner. Thus, the transfer hatch may be formed in the wall of the printer housing. Preferably, the transfer hatch is aseptically connectable to a sterile transfer bag allowing for the printing platform containing the three-dimensional printed object to maintain sterility during transfer of the printed object.

Optionally, the three-dimensional printer can comprise a membrane dispenser for dispensing a roll or sheets of membrane and which are printable on by means of the printing assembly. By means of the at least one printing head the membranes can be printed with proteins, antibodies, molecules, structural scaffolding or other printing materials. These printing materials might be stored in a tank located in the corresponding printing head within the sterile printer housing or might be provided from an external printing material source via a fluid line. The fluid line can be permanent fixed to the belonging printing head, particularly to keep the printing material and/or the printing head sterile. Alternatively, the fluid line can be removably connected to the belonging printing head, e.g. by a sterile connector.

The membrane dispenser can be driven hydraulically, pneumatically or magnetically. In order to be able to manufacture membrane of different sizes and shapes, the three-dimensional printer might further comprise a membrane cutter for cutting the roll or sheets of membrane into strips, sections, shapes, or pieces. The membrane cutter might be passive or active. A passive membrane cutter can comprise one or more fixed blade(s), thus utilizing the dispensing of the membrane roll to cut the membrane into strips utilizing the static blade(s) or a cutting die. An active membrane cutter can comprise one or more movable or rotatable blade(s), and thus, utilizing a mechanical motion of the blade(s) to cut the membrane, wherein the blades might be driven by a hydraulic/pneumatic actuator or an external magnetic drive mechanism.

Optionally, the three-dimensional printer further comprises a collection device for collecting the strips, sections, shapes, or pieces cut from the membrane, wherein the collection device can be moveable along the three axis framework in order to collect the strips etc. at different locations within the printer housing.

Depending on the printing materials, which are printed by means of the at least one printing head, it is required to support the fixation of the printing material. E.g. it might be required for printing material dissolved in aqueous solutions to support the drying process by providing dry air and/or hot air and thermoplastic material might require a cooling, particularly by blowing cool air to the printed material. Therefore, the three-dimensional printer might comprise a drying device for drying the three-dimensional printed object. Particularly, the drying device can comprise a dry air vent for providing air from an external air supply device. As an option, the dry air vent can comprise a sterilizing grade vent filter for filtering the incoming air to keep the interior of the printer housing sterile.

The hot air or cool air may be provided by an external device and flows through the vent into the interior of the printer housing, particularly to the printing platform. The three-dimensional printer may comprise a dispersal mechanism to evenly distribute the hot air or the cold air over the area of the printing platform, especially over the membrane, in order to dry the printed material onto the membrane. The three-dimensional printer may alternatively or additionally comprise a heating or cooling device being a disposable single-use device attached to the printer housing or included by the printer housing.

In order to keep a more or less constant pressure inside the printer housing, the excessive air from the dry air vent might be released into the exterior of the printer housing by means of a vent, particularly, a sterile vent. Correspondingly, waste air from the pneumatic actuators may be released into the printer body housing which is maintained at ambient pressure using the vent. The vent may also act as an air pressure regulating device keeping the interior of the printer housing at ambient pressure or at a predetermined underpressure or overpressure.

In order to keep the temperature inside the printer housing more or less constant at least a part of the printer housing can be formed as a thermal insulation or jacketing. The thermal insulation or jacketing can contain thermal barrier layers to prevent the transfer of heat or comprise tubing or capillaries for pumping a fluid through the insulation for the purposes of maintaining a constant temperature within the three-dimensional printer.

The three-dimensional printer may also comprise a temperature regulation device for maintaining a constant temperature within the printer housing. A temperature regulating device can measure the internal temperature of the three-dimensional printing assembly inside the printer housing and either heat or cool a fluid flowing into the printer housing via the in the printer housing to provide temperature regulation and/or incubation of the printed material.

The three-dimensional printer may also comprise an external temperature regulation device, such as a stainless steel container assembly, for maintaining a constant temperature within the printer housing. An external temperature regulating device may regulate the temperature by recirculating a temperature-controlled fluid inside of a jacketed container. The external temperature regulation device may alternatively utilize temperature controlled plates to provide a temperature exchange with the surface area of the printer assembly. The external temperature regulation device may utilize temperature measurements internal to the printer assembly for temperature regulation control.

The printing platform may be formed as the printing tray or may include the tray, e.g. positioned thereon, which is fillable with a liquid, preferably a nutrient rich liquid, for supplying living cells with an environment for growth during the printing process. The fluid in the tray can be recirculated, mixed, filtered, or drained and replaced according to the requirements of the printed structure.

Optionally an electric charge is providable by the printing platform or at least one printing head. Therefore, each of the at least one printing head(s) and/or the printing platform can comprise an electrode, in order to produce and distribute the small electric charge. A regulated electric charge can be utilized to stimulate cells located on the printing platform, particularly in a tray, during organ growth, incubation, or development.

Optionally, the three-dimensional printer may comprise a leveling device for horizontal leveling of the printing platform. By means of the leveling device it can be ensured that the three-dimensional printer and its printing platform or tray are leveled during the printing process. The leveling device can additionally be used to automatically calibrate the distance between the printing platform and the printing head in use.

The invention also relates to a printing system with a three-dimensional printing device comprising a sterilizable printer assembly including at least one printing head, a printing platform, and a driving mechanism adapted to achieve a relative displacement between the at least one printing head and the printing platform along two or three degrees of freedom.

A printer housing encloses the printer assembly in a sterile manner, and at least one aseptic connector fluidly connected to a corresponding one of the at least one printing head. The printing system also has a control device comprising a complementary driving mechanism adapted to drive a corresponding one of the driving mechanism of the three-dimensional printing device, at least one sterile printing material container fluidly connected with a corresponding one of the at least one aseptic connector and a controller for controlling the movement of the at least one printing head by means of the complementary driving mechanism and for controlling the ejection of the printing material by means of the at least one printing head.

Optionally, the complementary driving mechanism for moving the at least one printing head comprises a complementary magnetic actuator.

Optionally, the complementary magnetic actuator comprises an external motor driving a shaft to which a plurality of magnets is attached. It has to be understood, that a plurality of complementary magnetic actuators may be provided for each degree of freedom or movable axis. Each complementary magnetic actuator may utilize an external motor, preferably a stepper motor, containing a shaft and head with a plurality of magnets. The magnets can be ferrous magnets (such as iron), rare earth magnets (such as Neodymium), superconducting magnets, or magnetic fluids (ferrofluids). At least one magnetic head can mate directly or indirectly with an internal magnetic head of the magnetic actuator being internal and containing a plurality of magnets utilizing a bearing system or rigid wall separation. When the external motor rotates the magnetic connection between the external or internal magnets turns the internal magnetic head which in turn drives the corresponding printing head relative to the printing platform along one degrees of freedom. The driving can be performed by an internal threaded screw which is utilized to precisely move the printing platform along a track and or threaded screw within a three axis framework. This movement of the printing platform tray can be measured precisely with a tracking system. The system can auto-correct the position the printing platform by movement of the external magnetic drive mechanism to the exact coordinates required for the deposition of material by the printing device.

Optionally, the complementary driving mechanism for moving the at least one printing head comprises a controllable pneumatic source. The controllable pneumatic source is preferably an automated integrity testing device. In order to provide only a single controllable pneumatic source, this pneumatic source can be connected to a pneumatic manifold or a pneumatic multiplexer in order to sequentially fluidly connect a single one of a plurality of complementary driving mechanisms to the controllable pneumatic source. The three-dimensional printer, wherein an automated integrity testing device, preferably a Sartocheck® bag integrity testing device with a fine resolution pressure transducer, can be utilized as a measuring and pressure source for the pneumatic driving mechanism, such as a bag or bladder inflation mechanism, for positioning the printing platform/tray to the exact coordinates required for the deposition of printing material by the at least one printing head.

The manifold device linking the tubing from the three-dimensional printer to the measured pressure source can be controlled by the automated integrity testing device or an external device. The manifold device may link the tubing from the three-dimensional printer to the measured pressure source, which can be controlled by the automated integrity testing device or an external device. An electronic control section of manifold device which can be re-used can be externally attached to the sterile section of the manifold device which can be single-use.

Optionally, the printing system comprises a position tracking system, which is capable to determine the position of the printing platform relative to the at least one printing head along each of the three degrees of freedom. The position tracking system is connected to the controller for correcting position of the printing platform to the predetermined coordinates required for the deposition of the printing material by the at least one printing head. The position tracking system is capable to determine the position of the printing platform relative to the at least one printing head along each of the three degrees of freedom. The controller of the printing system can control the driving mechanism, e.g. auto-correct the inflation of the bags, in order to position the printing platform to the exact coordinates required for the deposition of printing material by the printing head.

Optionally, the printing system's three-dimensional printer further comprises a membrane cutter for cutting the roll or sheets of membrane into strips, sections, shapes, or pieces driven by a hydraulic/pneumatic actuator or an external magnetic drive mechanism controlled by the controller.

Optionally, the printing system's three-dimensional printer further comprises a drying device for drying the three-dimensional printed object, wherein the drying device includes an air supply device located outside the printer housing providing air through a vent in the printer housing towards the printing platform. The printing material can be dried utilizing a forced hot air or forced cold air provided by the air supply device through the vent, preferably through a sterilizing grade vent filter for filtering the air flowing into the printer housing. A heating or cooling device can take hot or cold air processed from an external device and can input it into the three-dimensional printing assembly. The heating or cooling device can contain a dispersal mechanism to evenly distribute the hot or cold air over the printing platform or a printed object, such as a membrane, to dry the printed material, e.g. onto the membrane. The heating or cooling device can be a disposable single-use device.

Optionally, the aseptic connector of the three-dimensional printer is fluidly connected to a feed or processing source including at least one of a bioreactor, a fermenter, a filtration train, a cross flow assembly, a membrane adsorber, a column, a centrifugation apparatus, a continuous centrifugation apparatus, an incubator, or other bioprocessing assemblies. The filtration train may include depth filters, pre-filters, sterilizing grade filters, ultra-filters, virus filters, etc. The cross flow assembly may include microfiltration or ultra-filtration cassettes.

The invention further relates to a printing method including the step of providing a three-dimensional printing device comprising a sterilizable printer assembly including at least one printing head, a printing platform, and a driving mechanism adapted to achieve a relative displacement between the at least one printing head and the printing platform along two or three degrees of freedom. A printer housing encloses the printer assembly in a sterile manner, and at least one aseptic connector fluidly connected to a corresponding one of the at least one printing head. The method proceeds by connecting at least one sterile printing material container fluidly with a corresponding one of the at least one aseptic connector, moving the at least one printing head by means of a complementary driving mechanism coupled to the driving mechanism and ejecting printing material by means of the at least one printing head for printing the printing material onto the printing platform or onto an object located on the printing platform under sterile conditions.

In case the printer assembly is not in a sterile condition that printing method may including the step of sterilizing the printer assembly. The sterilization may be carried out by using gamma-irradiation, autoclaving, or a chemical sterilant such as ethylene oxide or vaporized hydrogen peroxide.

In case the printing head is not pre-assembled with the sterilized printer assembly, a printing head assembly that is pre-sterilized, potentially by an alternate sterilization method than is used with the printer assembly, may be aseptically connected to the sterile printer assembly. A non-sterile printing head assembly may be inserted into a connection chamber and sterilized utilizing a chemical sterilant method such as ethylene oxide or vaporized hydrogen peroxide prior to removing the barrier between the printing head assembly and the printer assembly and prior to usage within the printer assembly.

One aspect of the invention relates to a manufacturing device that comprises a robotic arm assembly. The robotic arm assembly is internal to a sterilizable chamber and may comprise at least one actuator, at least one articulating support, at least one gripping assembly, and a driving mechanism adapted to achieve a relative displacement within the sterilizable chamber. The robotic arm assembly may be utilized for moving components and manufactured products from one place to another within a sterile chamber and/or from one sterile chamber to another in a connected network of sterile chambers. The robotic arm assembly may additionally be utilized for stacking and storing items as well as performing assembly of individual components.

The robotic arm assembly and the internal components making up the assembly, including the plurality of actuators, supports, articulations, seals, and plastic covering materials are sterile or sterilizable, e.g. by gamma-irradiation, autoclaving, or chemical sterilant such as ethylene oxide or vaporized hydrogen peroxide.

Optionally, the robotic arm assembly may comprise at least one pneumatic/hydraulic actuator comprising at least one of linear actuators, rolling diaphragm actuators, sterilizable linear actuators, sterilizable syringes, compressible cylinder, hydraulic rotary actuator, and hydraulic rotary vane actuators that are extendible in at least one direction by providing a fluid pressure to the inside of the actuating unit. The actuators apply force to an articulating support which extends or retracts the movement of a robotic arm. A plurality of actuators may be assembled with a plurality of articulating supports to provide the desired movements within the sterile chamber.

Optionally, the robotic arm assembly may move along a plurality of tracks using a wheeled and/or track assembly to increase the range of movement within the sterile chamber. The wheeled and/or track assembly may be powered by a hydraulic motor, a pneumatic motor, an electric motor, and/or a magnetic motor.

Optionally, the robotic arm assembly may comprise a soft robotic assembly. The soft robotic assembly may be actuated when pressurized with hydraulic and/or pneumatic pressure. A soft robotic assembly of sufficient complexity may allow the robotic arm to articulate around a fixed location instead of requiring a rotatable platform to direct the robotic arm in a particular direction.

Optionally, the robotic arm assembly may comprise a plurality of inflatable assemblies, where the supports of the robotic arm assembly comprise inflatable sections. The inflatable sections may comprise bags, bladders, and/or expandable reservoirs. The inflatable assemblies may be articulated along a seam and/or seal between two or more inflatable sections. The articulation of the robotic arm may be controlled by the movement of a plurality of cables connected to the inflatable sections.

Optionally, the fluid connections for control of the robotic arm assembly may be internal or external to the sterile chamber containing the robotic arm assembly. For an externally connected assembly the fluid lines for hydraulic/pneumatic control of the robotic arm assembly may be aseptically connected to the sterile chamber. The control mechanism for the robotic arm assembly may be manually controlled by an operator and/or automatically controlled utilizing a computer controlled setup. The computer-controlled setup may follow positional commands from a pre-programmed sequence stored in a memory storage unit.

Optionally, the robotic arm assembly may utilize a positional system for determining the distances and orientations of objects to determine how best to grab the objects and move them within the three-dimensional space of the sterile chamber. The positional system may comprise a plurality of cameras located on the sterile chamber and visual targets placed onto the components, items, and/or physical barriers. The visual targets may comprise variable augmented reality markers. Alternatively, a plurality of cameras may be placed onto the robotic arm assembly itself to position the robotic arm assembly. Alternatively, for additional precision a laser tracking system may be utilized externally by mounting it on the sterile chamber or internally by mounting it onto the robotic arm assembly. The robotic arm assembly may utilize other positioning techniques such as depth scanning, LIDAR, ultrasound, acoustic tracking, RFID/NFC tags, and other electronic methods to determine the position of the robotic arm operating within the three-dimensional space of the sterile chamber.

One aspect of the invention relates to a manufacturing device that comprises a vacuum-forming unit. The vacuum-forming unit assembly is internal to a sterilizable chamber and may comprise at least one rigid support structure, at least one vacuum platform with a plurality of holes, at least one vacuum pressure source, at least one heating element, at least one dispensing mechanism for dispensing moldable plastic sheets, at least one mold, and at least one vent filter assembly. The vacuum-forming unit assembly is intended for use in the manufacture of items with reproducible shapes by the forming of heated plastic sheet over a mold on a vacuum platform under vacuum pressure. Exemplarily, this manufacturing method may be used to produce a plurality of items with the same shape.

Optionally, the vacuum-forming unit assembly may utilize a vacuum pressure source which is external to the sterilizable chamber and is aseptically connected to such a vacuum source and/or is connected to a sterilizing grade air filter which is sized to sufficient capacity for the maximum vacuum pressure required for the forming process.

Optionally, the vacuum-forming unit assembly may utilize a vent filtration device which is sized to sufficient capacity to prevent the collapse of the sterile chamber under maximum vacuum pressure conditions.

Optionally, the vacuum-forming unit assembly may utilize a heating element for the heating of a deformable plastic sheet. The heating element heats a plastic sheet to make it easily deformable to take on the shape and structure of a fixed mold when it comes in contact with the mold under vacuum pressure. The heating element may be an electric heater, a single-use chemical heater device such as a device exploiting the exothermic oxidation of iron when exposed to air, a re-usable chemical heater device such as a device exploiting the exothermic crystallization of supersaturated solutions, or through the circulation of an externally-heated fluid source such as heated sterile filtered water, glycol, and/or steam. An externally-connected heating source may utilize an aseptic connection to connect to the heating element assembly within the sterile chamber.

Optionally, the sterilizable chamber may utilize a thermal barrier insulation to prevent structural damage to the sterile chamber and other internal components through the use of a heating element used with a manufacturing process device located within the sterile chamber.

Optionally, the vacuum-forming unit assembly may utilize a plurality of blades and/or die-cutting tools to cut the molded plastic sheet around the mold and to separate the desired formed product from the remaining plastic sheet.

One aspect of the invention relates to a manufacturing device that comprises an injection-molding unit. The injection-molding unit assembly is internal to a sterilizable chamber and may comprise at least one rigid support structure, at least one reservoir with meltable material, at least one heating element, at least one compression source, at least one dispensing unit, at least one solid mold comprising of at least two parts, and at least one vent filter assembly. The injection-molding unit assembly is intended for use in the manufacture of items with reproducible shapes by the melting of heated plastic material injected into a solid mold. Exemplarily, this manufacturing method may be used to produce a plurality of items with the same shape.

Optionally, the injection-molding unit assembly may utilize a heating element for the heating of moldable material, such as plastic pellets. The heating element heats the plastic pellets until they are melted within a chamber and the melted plastic is extruded into a solid mold made from at least two parts. The heating element may be an electric heater, a single-use chemical heater device such as a device exploiting the exothermic oxidation of iron when exposed to air, a re-usable chemical heater device such as a device exploiting the exothermic crystallization of supersaturated solutions, or through the circulation of an externally-heated fluid source such as heated sterile filtered water, glycol, and/or steam. An externally-connected heating source may utilize an aseptic connection to connect to the heating element assembly within the sterile chamber.

Optionally, the injection-molding unit assembly may utilize an external connection to supply the moldable material reservoir. The external connection for the moldable material supply into the sterile chamber may utilize an aseptic connection.

One aspect of the invention relates to a manufacturing device that comprises a laser-cutting unit. The laser-cutting unit assembly is internal to a sterilizable chamber and may comprise at least one rigid support structure, at least one two-axis positioning assembly, at least one laser assembly, at least one power source, and at least one substrate. The laser-cutting unit assembly is intended for use in the manufacture of custom-shaped items by cutting through a material substrate with a precision directed laser.

In case the laser-cutting device is not pre-assembled with the sterilized chamber, a laser-cutting device that is pre-sterilized, potentially by an alternate sterilization method than the sterilized chamber, may be aseptically connected to the laser-cutting assembly within the sterilized chamber. A non-sterile laser-cutting device may be inserted into a connection chamber and sterilized utilizing a chemical sterilant method such as ethylene oxide or vaporized hydrogen peroxide prior to removing the barrier between the printing head assembly and the printer assembly prior to usage within the sterilized chamber.

Optionally, the laser-cutting unit assembly may utilize an internal laser assembly which is powered by an external electrical connection, an internal battery storage mechanism, an inductively charged electrical connection, a chemical reaction, a microwave or visual line-of-sight power source, or other wireless power source. A visual line-of-sight power source is similar to a laser but does not require safety glasses to protect human eyes during operation since it is emitted within the visual part of the spectrum. Such a system to power local electronic devices is outlined in US patent application: US 2007/0019693 A1.

Optionally, the laser-cutting unit assembly may utilize a two-axis controller positional system, which may be driven by hydraulic, pneumatic, electric, or magnetically controlled motors. The positional system may utilize a system comparable to the system utilized for the three-dimensional printing control system.

Optionally, the laser-cutting unit assembly may be utilized for the laser labeling and/or barcoding of some materials or manufactured components.

Optionally, the sterile chamber may utilize a plurality of vent filters to remove heated air within the sterile chamber and to replace it a temperature regulated air. The airflow speed and number of air changes in the sterile chamber may serve as a temperature regulation mechanism. An external heat exchanger may be utilized to recirculate the air and control the temperature of the air entering into the sterile chamber.

Optionally, the sterile chamber may utilize a plurality of vent filters to remove aerosolized particulates or smoke from the sterile chamber. This may be utilized to remove aerosolized bioactive materials during the laser-cutting process, aerosols from spray coating, aerosolized particulates from CNC removal of materials, smoke or particulates from laser cutting, or general aerosolized particulates from manufacturing processes or material movements.

Optionally, the sterile chamber may utilize an absorptive material in the sterile chamber walls to block and/or reduce the intensity of the laser at the specific wavelengths that the laser assembly operates at. This will reduce the risk of exposure to operators who may or may not be wearing sufficient eye protection.

One aspect of the invention relates to a manufacturing device that comprises an ultrasonic-welding unit. The ultrasonic-welding unit assembly is internal to a sterilizable chamber and may comprise at least one rigid support structure, at least one ultrasonic welder comprising of a piston, a transducer, a converter, a booster, a sonotrode, a horn, an anvil, and at least two materials to weld together.

Optionally, the ultrasonic-welding unit assembly may be powered by an external electrical connection, an internal battery storage mechanism, an inductively charged electrical connection, a chemical reaction, a microwave or visual line-of-sight power source, or other wireless power source.

One aspect of the invention relates to a sampling system that comprises a single-use sampling system. The sampling system may be part of a manufacturing device and the sampling procedure may be part of a manufacturing operation. The single-use sampling system may comprise a collection container for holding a fluid sample, a fluid connection to the collection container, a single-use aseptic connection to a sterile chamber, a sampling method for drawing fluid material from the sterile chamber, and an aseptic disconnection method for removal of the container containing the fluid sample. The aseptically disconnected container may undergo measurement by an external measurement device, be used for storage of the material, or be used as a reference for the generation of other materials.

One aspect of the invention relates to a sampling system that comprises an external continuous sampling system. The external continuous sampling system may comprise an aseptic connection to a sterile chamber, a sampling method for drawing fluid material from the sterile chamber, an external measurement device, and a fluid connection to the external measurement device. The external measurement device may be a multi-use device which may additionally use single-use sensors for the testing of material from the sterile chamber. The fluid material after sampling or measurement may be aseptically returned to the sterile chamber via an aseptic connection or may be discarded through storage in a container or through a drain.

One aspect of the invention relates to a sampling system that comprises an internal sampling system. The internal sampling system may comprise a fluid connection to a collection container, a sampling method for drawing fluid material from the sterile chamber, and an internal measurement device. The internal measurement device may be integrated into the collection container and/or may be integrated into the sterile chamber.

One aspect of the invention relates to an optical measurement device. The optical measurement device, such as a microscopic camera, may contain a plurality of lenses, a zoom lens, an autofocus, and internal and/or external LED lighting.

Optionally, the optical measurement device may be inserted into a fixed position port within the sterile chamber using the same methods as used with the printing head insertion. The optical measurement device may be single-use or multi-use. A non-sterile optical measurement device may be connected to the sterile chamber and sterilized in place utilizing chemical sterilization methods. Alternatively, a pre-sterilized optical measurement device may be connected and inserted into the sterile chamber through an aseptic connection method.

Optionally, the optical measurement device may utilize the array of cameras utilized for the positioning system. The array of cameras may be positioned externally to the sterile chamber and view the internal assembly through a plurality of transparent windows and/or transparent material.

Optionally, the optical measurement device may optically examine, measure, record, and store the optical data in a digital storage device for comparison, trending, or time-lapse. The optical measurements may also be utilized to determine the profile, density, coverage, adherence, invasiveness, health, and viability of cell growth onto a structural support. The optical measurements may additionally examine other factors like color change from a chromic die for measuring pH, temperature, or other factors. The optical measurements may additionally be able to examine the contents for potential contamination of bacteria, fungi, viruses, or other unwanted cell growth.

One aspect of the invention relates to a chemical temperature-regulation device. The chemical temperature-regulation device of the sterile chamber may house a chemical reaction that comprises exothermic and/or endothermic reactions. The chemical temperature-regulation device may be internal to the sterile chamber and may be single-use. A single-use chemical temperature-regulation device may be endothermic such as by using ammonium nitrate or exothermic such as by exothermic oxidation of iron in air. It is possible for the chemical reaction to be recharged such as by exothermic crystallization of supersaturated solutions with sodium acetate. The chemical temperature regulation device may be sterilized along with the single-use chamber and discarded when the use of the chamber is completed.

One aspect of the invention relates to the aseptic connection of at least two sterile chambers together to form a network of sterile chambers where each sterile chamber performs a specific task. The network of sterile chambers increases the functionality of the items manufactured within the sterile chambers and the bioactive products which are derived from that manufacturing. The network of sterile chambers additionally allows the bioactive products to be printed within custom manufactured containers, supplied with nutrient rich media, undergo precise metered dosing of at least one drug product, undergo incubation within a temperature regulated environment, undergo analysis by an optical measuring device, undergo sampling by an aseptic sampling device and be aseptically removed from the network of chambers as a final manufactured bioactive product. The types of usages for such a network of sterile chambers include but are not limited to in vitro testing, the manufacture of biologically active cell products, the manufacture of implantable biological products, the manufacture of biologically active products on biosensors, the manufacture of biologically active products on diagnostic membranes, and custom manufactured filters coated with bioactive materials such as antibodies for use in capture or capture and elute processing methods. In vitro testing inside a network of sterile chambers includes the manufacturing of custom multi-well plates and coatings for in vitro testing, the use of multi-well plates for the screening of three-dimensional cell products, in vitro efficacy and toxicity studies of the effects of metered drug products on three-dimensional printed cells within multi-well plates. The manufacture of biologically active cell products inside a network of sterile chambers includes the printing of scaffolding materials for coating with bioactive products, the manufacture of complex multi-material assemblies for coating with bioactive products, the manufacture of complex multi-material medical devices with bioactive coatings, the printing of cells and cell products onto complex multi-material assemblies, the printing of cell products, cellular structures, scaffoldings, organs, and organ simulants grown from an individual patient's cells in a single-use bioreactor, processed, dispensed, incubated and grown in a single-use sterile environment. The manufacture of biologically active products on biosensors inside a network of sterile chambers includes the printing of biologically active products onto electronic devices and/or substrates to add to electronic devices for the detection of an analyte. The analyte may be detected through the combination of a biological component with a physicochemical detector. The manufacture of biologically active products on diagnostic membranes inside a network of sterile chambers includes the printing of biologically active products onto a diagnostic membrane and the assembly of the protective covering and/or device delivery tool for the diagnostic testing, reading of results, and test analysis. The manufacture of custom filters with biologically active products inside a network of sterile chambers includes the printing of biologically active products onto filter membranes which are assembled into a completed filtration unit. Bioactive materials such as antibodies may be utilized with a custom filter membrane for the capture and removal or for the capture and elution of a specific material from a filtered fluid.

Additional objects, advantages and features of the present invention will now be described in greater detail, by way of example, with reference to preferred embodiments depicted in the drawings.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

FIG. 1shows a three-dimensional printing device1comprising a printer housing5enclosing a printer assembly3. The inside of the printer housing5, particularly the printer assembly3, is sterilizable and disposable, i.e. intended for single use. The printer housing5can be formed of rigid walls or flexible walls7held open by a rigid internal and/or external skeleton.

The three-dimensional printing device1, particularly the printer assembly3and/or the inside of the printer housing5, can be sterilized by gamma-irradiation, autoclaving, or chemical sterilant (such as ethylene oxide or vaporized hydrogen peroxide). The electronic and controlling components for controlling the printer assembly3that are reused or that are sensitive to the sterilization method can be arranged outside the printer housing5and might be removably attached to the sterilized three-dimensional printer1during setup.

The at least one printing head of the three-dimensional printing device can positioned along three grades of freedom, such as the three axes x, y, and z, by means of a pressurized fluid, which can be provided by a fluid source. As an example, compressed air might be used as pressurized fluid. A regulated fluid source, which drives a three axis controller for the three-dimensional printer can be controlled by an automated integrity testing device12which takes air pressure from an incoming source and uses a sensitive, calibrated pressure transducer to accurately measure and dispense a precise pressure of fluid, such as air, to an outlet connection16.

The outlet connection16can be connected to a sterilizing grade filter20which feeds an electronically controlled tubing manifold22. The tubing manifold physically open and closes the connections to all of the air pressure tubing lines26which feed the three-dimensional printer1and is controlled by an electronic connection24to the controlling device12. The tubing manifold22and vent filter20unit can be sterilizable along with the three-dimensional printer1as a single piece and have an external electronically controlled device (not shown) that can be attachable to the tubing manifold22to control the opening and closing of each of the air pressure tubing lines26. The tubing lines26can deliver fluid pressure and be individually attached to fluid actuator, such as air actuators or inflatable bags or bladders, which can be utilized to move a printing tray32on a three axis framework30. The tubing manifold22can also individually vent each of the tubing lines26to remove air pressure in the inflatable bag/bladder or the air actuators. The three-dimensional printer is vented by a sterilizing grade air filter28so the internal pressure is always maintained at ambient. The printing tray32is push or pulled along the three axis framework30which can consist of threaded screws or tracks by the fluid actuators.

The printing tray32can be a flat platform or have walls which can hold a fluid during the printing process. The three-dimensional printer1can feature a plurality of fixed or movable printing heads with different functionality. In this embodiment the printing heads are fixed and features a spray deposition printing head36for coating the three-dimensional printed object with proteins, chemicals or molecules, a solid extruder printing head38for deposition of cells and other materials from a bioreactor, and a heated solid extruder head40for the deposition of structural elements. After the printing onto the tray has been completed the tray and printed structure can be removed via a transfer hatch34on the three-dimensional printer1wall. A sterile transfer bag (not shown) can be connected to the transfer hatch34where the tray and three-dimensional printed object can be removed and maintained within a sterile environment. The entire three-dimensional printer assembly1can also be placed in an incubator if further printing steps are required with the same unit.

FIGS. 2A-2Dshow an embodiment of inflatable bags or bladders utilized in a three-dimensional printer for the purpose of moving a platform on a three-axis framework.

FIG. 2Ais a top view of a two dimensional bladder50which can be inflated and vented out of a port52.

FIG. 2Bis a side view of a three-dimensional bag54which can be inflated and vented out of a port56.

FIG. 2Cis a side view of an assembly of three-dimensional bags60which are in a deflated state.

FIG. 2Dis a side view of an assembly of three-dimensional bags62which are in an inflated state.

The individual three-dimensional bags in the assembly go from larger sizes to smaller sizes and provide coarse to fine resolution for movement of a three-dimensional printing platform. The bags64and66provide the coarse resolution for inflation while bags68and70provide fine resolution for inflation. These are in place to move the three-dimensional printing platform into the correct position for the accurate deposition of printing materials within the specified coordinates.

FIGS. 3A-3Fshow an embodiment of the movements of the x-axis and z-axis controls of the three-dimensional printing platform utilizing inflatable bags or bladders.

FIG. 3Ais a top view of an embodiment of a three-dimensional printing device having a printable space100where a three-dimensional printing platform102in the following also called printing tray102, as a preferred printing platform, is pushed into a precise position by the coarse and fine resolution of the inflatable bags or bladders. In this case the three-dimensional printing tray102is in a centralized position and the bags104and106are in a deflated position.

FIG. 3Bis a top view of the printable space100where a three-dimensional printing tray102′, as a preferred printing platform, is pushed to a precise position to the left by the inflation of bag assembly106′. Bag assembly104remains in a deflated position.

FIG. 3Cis a top view of an embodiment of the printable space100where a three-dimensional printing tray102″, as a preferred printing platform, is pushed to a precise position to the right by the inflation of bag assembly104′ and the deflation of bag assembly106.

FIG. 3Dis a top view of an embodiment of the printable space100where a three-dimensional printing tray102′″, as a preferred printing platform, is pushed to a precise position forward by the inflation of bag assembly108′. Bag assemblies104,106, and110remain in a deflated position.

FIG. 3Eis a top view of an embodiment of the printable space100where a three-dimensional printing tray102″″, as a preferred printing platform, is pushed to a precise position backward by the inflation of bag assembly110′ and the deflation of bag assembly108. Bag assemblies104and106remain in a deflated position.

FIG. 3Fis a top view of an embodiment of the printable space100where a three-dimensional printing tray102′″″, as a preferred printing platform, is pushed to a precise position in the forward right direction by the inflation of bag assembly108′ in the forward direction and bag assembly104′ in the right direction. Bag assemblies106and110are in a deflated position.

FIGS. 4A and 4Bshow an embodiment of the movements of the y-axis of the three-dimensional printing platform or printing tray124utilizing inflatable bags or bladders.

FIG. 4Ais a side view of an embodiment of a three-dimensional printer120where a three-dimensional printing tray124is pushed into a precise position by the coarse and fine resolution of the inflatable bags or bladders. In this case the three-dimensional printing tray124is in a centralized position and the bag assembly122is in a deflated position.

FIG. 4Bis a side view of an embodiment of a three-dimensional printer120where a three-dimensional printing platform or printing tray124′ is pushed upwards into a precise position by the inflation of bag assembly122′. The elevation of the tray by the inflation of bag assembly122′ also lifts the bag assemblies (not shown) utilized for the x-axis and z-axis controls on the three-axis framework.

FIGS. 5A and 5Bshow an embodiment of alternate mechanisms to drive the three-dimensional printing tray or printing platform into a precise location for deposition of printing material.

FIG. 5Ais a side view of an embodiment of an pneumatic actuator140which utilizes compressed air150through an airline144which converts the air pressure to a mechanical motion, utilizing a valve stem or a rotary actuator146, and driving an internal screw mechanism148which is attached to the three-axis framework, which in this embodiment comprises a threaded screw142. This mechanism allows the pneumatic actuator140to move in a forward or backwards direction154along the path of the threaded screw142, or alternatively a track. The waste compressed air is expelled out of outlet152and into the three-dimensional printer chamber where it is vented by an appropriately sized sterilizing grade vent filter (not shown) which maintains the chamber at ambient pressure.

FIG. 5Bis a side view of a magnetic motor control device which can drive an internal screw to move the three-dimensional printing tray. The magnetic motor control device comprises an external motor156, a shaft158and a linkage device160containing a plurality of magnets162. The linkage device160connects to a location on a three-dimensional printer body wall164which can be rigid or flexible. An internal linkage device168contains a plurality of magnets166which mates with the plurality of magnets162of the external linkage device160. The external motor156rotates the external linkage device160and magnets162, and thus, drives the internal magnets166and the linkage device168which results in a turning motion172of an internal threaded screw170. A friction reducing assembly, such as ball bearings (not shown), can be utilized on the internal and the external linkage devices160and168to reduce the friction from the drive magnets on the three-dimensional printer wall164. This turning motion172of the internal threaded screw170allows the three-dimensional printing tray or printing platform to move precisely in a forward or reverse direction along rotational axis of the internal threaded screw170.

FIGS. 6A-6Dshow an embodiment of a precision tracking system or position tracking system to verify the coordinate location of the printing platform also called printing tray along the three axis framework. The precision tracking system can additionally be utilized to calibrate the distance between the printing platform or the printing tray and the printing head in use. The precision tracking system can determine the distance and location of multiple points on the printing platform or the printing tray and make the adjustments to level the printing platform in line with the printing head. This function can be performed prior to printing and/or during the printing function.

FIG. 6Ais a front view of a three-dimensional printer180where a laser emitting device182sends out a signal186which is reflected by means of a plurality of mirrors184located on the printing platform or tray.

FIG. 6Bis a front view of the same three-dimensional printer180, as inFIG. 6Awhere a laser detection device188detects the angle and time the signal186′ takes to reach the detector giving precise coordinates of the three-dimensional printing tray location. This information is relayed to the controller where the printer can precisely deposit material onto the printing tray along the three axis framework.

FIG. 6Cis a front view of a three-dimensional printer180comprising a camera array190, wherein the camera array190observes a visual target192attached to the three-dimensional printing platform or formed with the three-dimensional printing platform, also called a printing tray. The location and size of the visual target192can provide precise coordinates of the three-dimensional printing tray location which is relayed to the controller where the printer can precisely deposit material onto the printing tray along the three axis framework.

FIG. 6Dis an embodiment of the visual target194which is recognized by the camera array190.

FIG. 7shows an embodiment of a single-use bioreactor and filtration assembly connected to a single use three-dimensional printer via an aseptic connector, wherein the three-dimensional printer is capable to form a printed object. This embodiment shows a gamma irradiated assembly containing a single-use bioreactor200which is connected to a filtration train via an aseptic connector202. The filtration train can comprise a plurality of filters including but not limited to a depth filter204, a pre-filter206, and a sterilizing grade filter208.

The filter train is connected to a surge vessel container214via an aseptic connector210. The surge vessel container214fills with the material filtered from the bioreactor, which can be driven by a constant pressure or constant flow source. A sterilizing grade vent filter218allows the surge vessel container214to vent during filling. After the filtration process is complete or the surge vessel container214is full the valve212to the filter train is closed and a regulated compressed air line216is attached to the sterilizing grade air filter218. The pressure drives the liquid up a dip tube and into a tubing piece which is connected via an aseptic connector220to the three-dimensional printing assembly230. The material from the surge vessel container214can be concentrated utilizing a cell retention and concentration device, preferably a gamma irradiatable Hydrocyclone (not shown). The processed material can be deposited onto the three-dimensional printing tray or printing platform224in a precise location through extrusion, spray deposition, or by mixing with a structural component through one or more of the at least one printing head of the printer assembly222. The three-dimensional printed object226on the three-dimensional printing tray224is formed by layer-by-layer additive printing of material from the printer assembly222. The three-dimensional printer230vessel, which can be rigid or flexible, is vented by a sterilizing grade vent filter228. After the printing of the three-dimensional printed object226has been completed the three-dimensional printing tray224and printed structure226can be removed via a transfer hatch232on the three-dimensional printer230wall. A sterile transfer bag234can be connected to the transfer hatch232via a sterile connection236. Thus, the printing tray224and three-dimensional printed object226can be removed from the three-dimensional printer230while being maintained within a sterile environment.

FIG. 8shows an embodiment of two single-use bioreactors and filtration assemblies connected to a single-use three-dimensional printer via aseptic connectors to form a printed object. This embodiment shows a gamma irradiated assembly containing two single-use bioreactors250and280which are connected to filtration trains via aseptic connectors252and282. The filtration trains can comprise a plurality of filters including but not limited to pre-filters254and284and sterilizing grade filters256and286. The filter trains can be connected to a surge vessel container262and292via aseptic connectors258and288. The surge vessel containers262and292fill with the material filtered from their respective bioreactors, which can be driven by a constant pressure or constant flow source. Sterilizing grade vent filters266and296allow for the surge vessel containers262and292to be vented during filling. After the filtration process is complete or the surge vessel containers262and292are full, the valves260and290to the filter trains are closed and regulated compressed air lines264and294are attached to the sterilizing grade air filters266and296. The pressurized air provided by the air lines264and294drives the liquid up a dip tube and into a tubing piece which is connected via aseptic connectors270and298to a single three-dimensional printing assembly300.

The material from the surge vessel containers262and292can be deposited onto the three-dimensional printing tray or printing platform304in a precise location through extrusion, spray deposition, or by mixing with a structural component through one or more of the printing heads on the printer assembly302. The three-dimensional printed object306on the three-dimensional printing tray304is formed by layer-by-layer additive printing from material coming from either bioreactor or from both in a specified mixture from the printer assembly302.

The three-dimensional printer300vessel, which can be rigid or flexible, is vented by a sterilizing grade vent filter308. After the printing of the three-dimensional printed object306has been completed the three-dimensional printing tray304and printed structure306can be removed via a transfer hatch310on the three-dimensional printer300wall.

FIG. 9shows an embodiment of a single-use bioreactor, a filtration assembly, and a cross flow assembly connected to a single-use three-dimensional printer via an aseptic connector to form a printed object. This embodiment comprises a gamma irradiated assembly containing a single-use bioreactor320which is connected to a filtration train via an aseptic connector322. The filtration train can comprise a plurality of filters including but not limited to a depth filter324, a pre-filter326, and a sterilizing grade filter328. The filter train is connected to a surge vessel container338via an aseptic connector330. The surge vessel container338fills with the material filtered from the bioreactor which can be driven by a constant pressure or constant flow source. A sterilizing grade vent filter334allows the surge vessel container338to vent during filling. After the filtration process is complete or the surge vessel container338is full, the valve336to the filter train is closed and a regulated compressed air line332is connected fluidly with the sterilizing grade air filter334. The pressure drives the liquid up a dip tube and into a tubing piece which is connected to a pre-sterilized cross flow assembly340.

The cross flow assembly340can comprise a plurality of microfiltration342and/or ultrafiltration cassettes344in varying sizes. The cross flow assembly is connected to a surge vessel container350via an aseptic connector346. The surge vessel container350fills with the material filtered and/or concentrated from the cross flow assembly which can be driven by a constant pressure or constant flow source. A sterilizing grade vent filter354allows the surge vessel container350to vent during filling. After the cross flow processing is complete or the surge vessel container350is full, the valve348to the cross flow assembly is closed and a regulated compressed air line352is attached to the sterilizing grade air filter354. The pressure drives the liquid up a dip tube and into a tubing piece which is connected via an aseptic connector356to the three-dimensional printing assembly360.

The material from the surge vessel container350can be deposited onto the three-dimensional printing tray or printing platform362in a precise location through extrusion, spray deposition, or by mixing with a structural component through one or more of the printing heads on the printer assembly358. Prior to gamma irradiation membrane and/or diagnostic strips364can be prepositioned onto the three-dimensional printing tray362. The three-dimensional printing assembly358can spray deposit proteins and/or other concentrated ultra-filtered materials onto the membranes strips for use in diagnostic analysis. Additionally other structural components can be added to the membrane strips by layer-by-layer additive printing of material from the printer assembly358. After the printing onto the three-dimensional printed membrane strips364has been completed the three-dimensional printing tray362and printed membrane strips364can be removed via a transfer hatch366on the three-dimensional printer360wall.

FIG. 10shows an embodiment of a single-use bioreactor, a centrifugation assembly, a filtration assembly, a membrane adsorber assembly, and a cross flow assembly connected to a single-use three-dimensional printer via an aseptic connector to form a printed object. This embodiment comprises a gamma irradiated assembly containing a single-use bioreactor370which is connected to a continuous flow centrifuge374via an aseptic connector372. The continuous flow centrifuge removes the heavy particulates from the bioreactor harvest and allows the supernatant to continue into the filtration train assembly. The filtration train can comprise a plurality of filters including but not limited to a depth filter (not shown), a pre-filter376, and a sterilizing grade filter378. The filter train is connected to a surge vessel container384via an aseptic connector380. The surge vessel container384fills with the material filtered from the bioreactor which can be driven by a constant pressure or constant flow source. A sterilizing grade vent filter388allows the surge vessel container384to vent during filling. After the filtration process is complete or the surge vessel container384is full, the valve382to the filter train is closed and a regulated compressed air line386is fluidly connected to the sterilizing grade air filter388. The pressure drives the liquid up a dip tube and into a tubing piece which is connected to a pre-sterilized membrane adsorber390. The membrane adsorber390is a chromatographic membrane carrying functional groups for the reversible binding of biomolecules. The desired molecules can be captured with the membrane adsorber and eluted at a later time or undesirable molecules can be removed by membrane adsorption before further processing. The membrane adsorber390can be connected to a pre-sterilized cross flow assembly392. The cross flow assembly392can comprise a plurality of microfiltration and/or ultrafiltration cassettes in varying sizes. The cross flow assembly is connected to a surge vessel container398via an aseptic connector394. The surge vessel container398fills with the material filtered and/or concentrated from the cross flow assembly392which can be driven by a constant pressure or constant flow source. A sterilizing grade vent filter412allows the surge vessel container398to vent during filling. After the cross flow processing is complete or the surge vessel container398is full, the valve396to the cross flow assembly is closed and a regulated compressed air line410is attached to the sterilizing grade air filter412. The pressure drives the liquid up a dip tube and into a tubing piece which is connected via an aseptic connector414to the three-dimensional printing assembly400.

The material from the surge vessel container398can be deposited onto the three-dimensional printing tray or printing platform404in a precise location through extrusion, spray deposition, or by mixing with a structural component through one or more of the printing heads on the printer assembly402. Prior to gamma irradiation membrane and/or diagnostic strips406can be pre-positioned onto the three-dimensional printing tray404. The three-dimensional printing assembly402can spray deposit proteins and/or other concentrated ultra-filtered materials onto the membranes strips for use in diagnostic analysis. Additionally other structural components can be added to the membrane strips by layer-by-layer additive printing of material from the printer assembly402. After the printing onto the three-dimensional printed membrane strips406has been completed the three-dimensional printing tray404and printed membrane strips406can be removed via a transfer hatch408on the three-dimensional printer400wall.

FIG. 11shows an embodiment of a single-use three-dimensional membrane printer with a three-dimensional axis framework for a stacking or storage tray. This embodiment shows a gamma irradiated membrane printing assembly500containing a membrane dispensing and print section502which has a membrane roll510suspended by a dowel (not shown). The membrane from the membrane roll510is dispensed utilizing a magnetic roller assembly512which mates to an external motor with a magnetic head (not shown) to drive the movement of the membrane through the membrane printing assembly500. The motor speed of the external motor controls the speed at which the magnetic roller assembly512moves and at which the membrane is dispensed from the roll510. An assembly of passive rollers514keeps the membrane straight and at tension as the membrane is dispensed. These passive rollers514can be present throughout the membrane printing assembly500to maintain tension and a straight path for the membrane.

Fluid material, which may be provided by a single-use bioreactor, enters the printing head518from the tubing516under pressure. The electronically controlled printing head518dispenses the fluid520onto the membrane with a specific pattern. The magnetic roller assembly512can reverse the membrane if multiple passes of fluid and/or structural deposition of material onto the membrane is required for the process. The printed membrane section then moves through an opening in a wall522where it enters the drying section of the assembly504where the membrane can undergo drying by utilizing heated or cooled sterile air.

A sterilizing grade vent filter524can be utilized to bring air that has been heated or cooled from an outside source. The air can enter into the membrane printing assembly500through a diffuser block526, wherein the diffuser block526takes the incoming air and diffuses it so that there is an even application of the heated or cooled air528across the membrane to allow for even drying. The membrane section then passes through an opening in a wall530to a cutting section506of the membrane printing assembly500. Both walls522and530serve as a physical barrier to prevent the overheating of components within the membrane assembly if heated air is used to dry the printed materials on the membrane.

In a simple embodiment the walls522and530can be a simple layer of thin plastic. In a more complex embodiment the walls522and530can contain a thermal insulation to prevent the transfer of heat to other areas of the assembly. In an even more complex embodiment the walls522and530can contain a capillary network of tubing which can be connected to a cooling or heated water source to prevent thermal transfer and offset the temperature of the air used to dry the printed membrane. The walls of the entire three-dimensional printing assembly can also be jacketed to maintain a constant desired temperature.

The cutting section506contains at least two movable clamps including one top clamp532and one bottom clamp534. The top clamp532and the bottom clamp534can clamp down onto the membrane in a specified section preferably one that has not undergone printing and allows for a rigid hold so that the membrane can be cut into sections by a knife cutting assembly536. A plurality of cutting knives or cutting dies in the knife cutting assembly536can cut the membrane into horizontal and/or vertical sections, or die-cut shapes to a specified sizing as required. The knife cutting assembly536can feature passive knives under the membrane which cuts the membrane into sized vertical strips as the membrane is fed through it. A horizontal cutting knife can be mechanically actuated to cut the membrane strips at a specific length. The top clamp532the bottom clamp534and an actuated horizontal cutting knife from the knife cutting assembly536can all be driven pneumatically from an external air pressure source or magnetically driven from an external mechanical source. As the membrane is cut by the knife cutting assembly536the membrane strips538falls into a collection tray or collection device540in the membrane strip collection section508of the assembly. This collection tray can move along a 3 axis framework542to stack the membrane strips538as they fall into the tray.

In this assembly multiple trays, as preferred collection device, can be utilized along the same three axis framework to increase the area of the membrane strips collected into the trays. The trays can also have a rotatable feature where the internal tray can be rotated along the holding platform to increase the area where the membrane strips538can be stacked. In other words the trays may have two linear degrees of freedom and one rotational degree of freedom. The number of trays and the sizing type of trays used can be determined by the size of the membrane roll, the size and the number of strips that need to be cut, and if the printed material on the membrane is sensitive and cannot undergo stacking or can only undergo limited stacking. The entire membrane printing assembly can be maintained at ambient pressure through a sterilizing grade vent filter544. After the printing process has completed and the membranes strips538are placed in the collection trays540the trays can be removed through the transfer hatch546. A sterile transfer bag not shown can be attached to the transfer hatch546if the maintenance of sterility is required.

FIG. 12illustrates an embodiment of a single-use three-dimensional printer with a multi-segmented tile printing platform and a membrane dispenser to form a plurality of printed filtration devices.

This embodiment shows a gamma irradiated filter device printing assembly650containing a membrane dispenser which has a membrane roll652suspended by a dowel (not shown) and only takes up a portion of the horizontal space within the printing assembly. The membrane from the membrane roll652is dispensed utilizing a magnetic roller assembly654which mates to an external motor with a magnetic head (not shown) to drive the movement of the membrane through the filter device printing assembly650. The motor speed of the external motor controls the speed at which the magnetic roller assembly654moves and at which the membrane is dispensed from the roll652. An assembly of passive rollers656,658, and660keeps the membrane straight and at tension as the membrane is dispensed. These passive rollers656,658, and660can be present throughout the filter device printing assembly650to maintain tension and a straight path for the membrane. In a more complex embodiment a pleating device (not shown) may be utilized to form pleats with the membrane, which are used to increase the surface area of the membrane available within a limited space.

The membrane sheet from membrane roll652is spooled out into a holder670where a cutting die662presses down utilizing the force from an internal shaft664from an externally mounted piston666to cut through the membrane. This forms a die-cut membrane shape674which falls through the holder670and maintains placement via guide bars672. The die-cut membrane674is guided to a partially formed filter device676which was printed by a three-dimensional printing head700by layer-by-layer additive printing of a plastic material, preferably polypropylene by a heated extrusion head, but could also be made of Polylactic acid or polylactide (PLA), Acrylonitrile butadiene styrene (ABS), or other printable material which can be fed to the printing head700as a spooled material702, heated and extruded onto the printing platform. The partially formed filter device676can include an opening for the fluid to pass through, material to form the body of the filter device, a cavity to fit the die cut membrane674, and structural material to assist in the building of overhanging structures such as a bridge. Biologic material originating from a bioreactor including proteins, ultra-filtered materials, or other materials can be spray deposited using a separate printing head onto the membrane prior to die-cutting or prior to the completion of the filter device.

After the die-cut membrane674is placed on the cavity in the partially formed filter device676the individual tiles704of the multi-segmented printing platform708can be moved utilizing magnetic roller assemblies710and712which mates to an external motor with a plurality of magnetic heads (not shown) to drive the movement of the individual tiles704via a series of rotating screws in a stepwise movement through the filter device printing assembly650which mimics a sliding tile puzzle. The multi-segmented printing platform708has an empty space706which allows for all individual tiles704to be moved to all possible positions on the printing platform. The individual tiles704containing the partially formed filter devices676containing the die-cut membranes674are moved in a stepwise movement to the printing head700which occupies a portion of the horizontal space not taken up by the membrane dispensing assembly. The printing head700then seals the die-cut membrane674by extruding material around the rim of the partially formed filter device676. The printing head700then prints the remainder of the filter device by layer-by-layer additive printing to form a completed filter device678containing a sealed membrane layer. In a more complex embodiment the die-cut membrane674may be ultrasonically welded onto the rim of the partially formed filter device676using an ultrasonic welding device (not shown) described inFIG. 32. The ultrasonic welding device (not shown) may additionally ultrasonically weld the at least one bottom part of a partially formed filter device676to the at least one top part of corresponding partially formed filter device (not shown) to form a completed filter device678. The ultrasonic welding device (not shown) may additionally ultrasonically weld a pleated section of membrane (not shown) together to form a pleated membrane assembly (not shown) prior to assembly into a pleated filter device (not shown). The completed filter device individual printing platform tile is moved into position in a stepwise movement to enter the completed filter device bin690. A rigid blade682cuts the bottom of the completed filter device680to remove it from the printing platform tile. A printed structural element, like a raft, could be utilized to ease the process of removing the completed filter device680with a clean break from the rest of the printing platform to reduce any potential defect from the cutting/removal process. A movable guide on a hinge684can push the completed filter device into the completed filter device bin690. This guide can go along the guide path686and push completed filter devices688throughout the bin690. The movable guide can be controlled externally by a magnetic motor assembly or an insertable shaft motor (not shown).

The waste components of the membrane roll after being die-cut can be spooled into a membrane waste bin692which can collect the remaining membrane694and make for easy disposal after the printing process has completed. The entire membrane printing assembly can be maintained at ambient pressure through a sterilizing grade vent filter714. After the printing process has completed the completed filter devices688located in the holding bin690can undergo sterile transfer using the transfer hatch716if the maintenance of sterility is required.

FIGS. 13A-13Fshow an embodiment of the connections steps to insert the printing head into the sterile three-dimensional assembly body.

FIG. 13Ais a side view of an embodiment that shows a gamma irradiated three-dimensional printer assembly body550, with a rigid or flexible wall, containing a printing head insertion assembly548containing a cap assembly552internal to three-dimensional printer assembly with an internal dip tube554. The external portion of the printing head insertion assembly548contains a removable cap560, a fluid inlet port556, and a fluid outlet port558.

FIG. 13Bis a side view of the removable cap560′ being removed from the printing head insertion assembly548and the electronically controlled printing head562being inserted into the assembly.

FIG. 13Cis a side view of the electronically controlled printing head562inserted into the printing head insertion assembly548and twisted one position into place564.

FIG. 13Dis a side view of the printing head insertion assembly548with a tubing piece566inserted into the fluid inlet port556where a chemical sterilant or sanitizer568fills the cap assembly552sterilizing the internal section of the electronically controlled printing head. The chemical sterilant or sanitizer568can comprise a liquid like 30% hydrogen peroxide, or may comprise a gas sterilant such as vaporized hydrogen peroxide or ethylene oxide.

FIG. 13Eis a side view of the printing head insertion assembly548with a tubing piece570inserted into the fluid outlet port558where the chemical sterilant or sanitizer is removed by vacuum pressure after the period of time required to sterilize the printing head is completed. The internal dip tube554can be utilized to remove a fluid chemical sterilant or sanitizer or condensate from a gas chemical sterilant.

FIG. 13Fis a side view of the printing head insertion assembly where the printing head is twisted572into the second position where the cap assembly552′ drops into the interior of the three-dimensional printer assembly body. The cap assembly552′ can be moved manually to a holding bin within the three-dimensional printer assembly body by tilting the three-dimensional printer assembly until the cap assembly552′ falls into place. The sterilized printing head562is ready to print within the three-dimensional printed assembly.

FIGS. 14A-14Eshow an embodiment of the setup process for a flexible wall three-dimensional printer from the packed shipping configuration.

FIG. 14Ais a side view of an embodiment that shows a gamma irradiated three-dimensional printer assembly600with flexible walls that is folded flat in a configuration for shipping. The entire assembly can be enclosed in multiple gamma irradiatable bags for ensuring sterility of the bags when moving from the receiving to the end use facility. The shipping configuration consists of the air bags completely deflated or the actuators and/or magnetic drivers in the minimal position. The bag handles602and604on the top of the three-dimensional printer assembly600are folded over. The printing head insertion assembly606rest inside of the printing platform tray during shipping. The sterilizing grade venting filter608is laid on its side and is capped off with cap610. A bag integrity testing device, preferably a Sartocheck® automated bag integrity testing unit, can be utilized by inserting the bag assembly between two plates containing a fleece layer to test the integrity of the bag seal and ensure there are no leaks prior to use.

FIG. 14Bis a side view of an embodiment that shows a gamma irradiated three-dimensional printer assembly600′ with the handles extended and the operator pulling the handles602′ and604′ upwards. The three-dimensional printing assembly bag chamber can be inflated with sterile air from an external source not shown. The folded screws or tracks forming the three axis framework and the internal support skeleton can be manually snapped into place by the operator by pulling up and pushing down on the external handles602′ and604′. The printing head insertion assembly606is supported by an internal crossbar support that can also be snapped into place by the operator. The cap610on the venting air filter608can be removed after the support skeleton is in place to ensure that the flexible walls do not collapse when vented to the outside.

FIG. 14Cis a side view of an embodiment that shows a threaded screw assembly620making up the three axis framework where the printing platform is precisely moved. The threaded screw assembly620is in a folded configuration for shipping where the first threaded screw section622is separated by the second threaded screw section624by a hinge626and an internal locking mechanism628.

FIG. 14Dis a side view of an embodiment that shows a threaded screw assembly620′ which is folded back into a vertical position using hinge626by the operator pulling back on the external handles to unfold the bag. The threaded screw assembly620′ is in a folded back where the second threaded screw section624′ is in a vertical position and is in line with the first threaded screw section622.

FIG. 14Eis a side view of an embodiment that shows a threaded screw assembly620″ where the second threaded screw section624′ is pushed down to mate with the first threaded screw section622covering the hinge assembly626and the locking mechanism while forming a single threaded screw body620″. The locking mechanism628clicks into place ensuring that the threaded screw cannot be detached unless the locking mechanism is disengaged. The operator pulls the external handles down to achieve the locking of the threaded screws for the three axis framework and/or other internal support elements forming the internal support structure for the three-dimensional printer assembly.

FIGS. 15A-15Dshow an embodiment of the movements of the x-axis, y-axis, and z-axis controls of the three-dimensional printing platform utilizing hydraulically-driven actuators.

FIG. 15Ais a side view of a basic embodiment of a manually controlled hydraulically driven three-dimensional printing device800having a sterile chamber where a three-dimensional printing platform814containing a printing tray824, as an exemplary printing platform, is pushed and/or pulled into a precise position by the hydraulic actuators810,812,816,820for three-dimensional printing by a plurality of printing heads838. In this basic embodiment the internal hydraulic actuators810,812,816,820are controlled and driven by an external set of manual actuators802. In more complex embodiments the manually-controlled external set of manual actuators802may be replaced by an automated computer-controlled setup (not shown). Some manual actuators802may be sized to hold a volume of fluid equivalent to the capacity of the corresponding internal hydraulic actuator, as is the case with hydraulic actuators816and820. Other manual actuators802may additionally be sized to hold a volume of fluid equivalent to or larger than the combined capacities of multiple corresponding internal hydraulic actuators, as is the case with actuators810and812, where multiple internal hydraulic actuators810and812are uniformly controlled by an individual external manual actuator802. The hydraulic actuators setup may be filled with an incompressible fluid, such as sterile filtered purified water, to drive the internal hydraulic actuators810,812,816,820. Hydraulically-driven actuators have the following advantages: they have a higher pressure-to-weight ratio over pneumatic actuators, they can hold force and torque constant without the pump supplying more fluid or pressure, they can produce a higher force with respect to a pneumatic cylinder of equal size, and they can be placed at a greater distance away from the device800with minimal loss of power. The disadvantage is the potential for leakage in the tubing lines within the setup. In this embodiment a sterile drain830, containing a sterilizing-grade filter832, is positioned in the bottom-most portion of the chamber to remove any potential leakage of hydraulic fluid from the setup. Specialty hydraulic fluid oils may be utilized for particular applications where high-weight transfers are required but their use in a clean, sterile environment would require additional precautions for tubing strength and secure connections to prevent the leakage of hydraulic fluids into the sterile chamber. A hydraulic actuator provides unidirectional force through a unidirectional stroke. Examples of hydraulic actuators for the following setup include but are not limited to linear actuators, rolling diaphragm actuators, sterilizable linear actuators, sterilizable syringes, hydraulic rotary actuator, and hydraulic rotary vane actuators. In this embodiment the three-dimensional printing platform814and the printing tray824are at the lowest height of the compressed y-axis hydraulic actuators. The hydraulically-driven three-dimensional printing device800utilizes an external set of manual actuators802to drive the internal hydraulic actuators810,812,816, and820. The hydraulic tubing lines are filled with a sterile filtered fluid, such as sterile filtered purified water, through a filter assembly804. The filter assembly804may contain a sterilizing-grade filter through which the fluid input is provided to fill the tubing lines806, hydraulic actuators802,810,812,816,820, and any fluid reservoirs (not shown). The filter assembly804may contain a ‘T’ connection with a valve to separate the incoming fluid line from the rest of the hydraulic assembly. A sterilizing-grade vent filter (not shown) may be employed to properly vent the tubing lines806, hydraulic actuators802,810,812,816,820, and fluid reservoirs (not shown) of any residual air during the filling procedure. The hydraulic tubing lines806enter into the sterile chamber through a port808in the chamber. The hydraulic tubing lines806connect to the y-axis controlling hydraulic actuators810and812, which vertically raise and lower the height of the printing platform814and printing tray824, to the x-axis controlling hydraulic actuator816, which moves the printing tray824horizontally, and to the z-axis hydraulic actuator820, which moves the printing tray horizontally and orthogonally to the direction in which the actuator816move the printing tray. In this embodiment the x-axis hydraulic actuator816is connected to a printing tray assembly822with a connection piece818that moves along on a track on the printing platform814. The z-axis hydraulic actuator820is connected to the printing tray assembly822and moves along with it while moving the printing tray824along a track on the printing tray assembly822. This precisely positions the printing tray824for three-dimensional printing through the dispensing of material from a plurality of printing heads838.

FIG. 15Bis a side view of a basic embodiment of a manually-controlled hydraulically driven three-dimensional printing device800. The external set of manual actuators802′ are manually depressed by an operator, which drives the internal hydraulic actuators to move into positions810′,812′,816′, and820′. The hydraulic fluid, which in this exemplary embodiment is sterile filtered purified water, enters into the chamber through tubing lines806and places pressure on the actuators causing the piston rods from the actuators to extend thus moving the positions of the 3-axis framework to position the printing tray for the plurality of printing heads838. In this embodiment the three-dimensional printing platform814and the printing tray824are at the highest height of the extended y-axis hydraulic actuators810′ and812′. In this embodiment a minor leakage of hydraulic fluid from an internal tubing line connection is removed by adding a vacuum assembly collection flask834and a vacuum pump836to the sterile drain assembly830. The vacuum assembly collection flask834is connected to the sterilizing-grade filter832on the sterile drain assembly830. The vacuum pump836is turned on and the hydraulic fluid material that leaked into the sterile chamber is removed and discarded from the vacuum assembly collection flask834. Additional hydraulic fluid may be added to the hydraulic fluid tubing line806using the filter assembly804. In more complex embodiments the manually-controlled external set of manual actuators802may be replaced by an automated computer-controlled setup (not shown).

FIG. 15Cis a top view of the hydraulically-driven three-dimensional printing device800where the printing tray824is moved along the x-axis and z-axis with corresponding hydraulically driven actuators816and820. In this embodiment the x-axis hydraulic actuator816is in a fixed position on the printing platform814. The x-axis hydraulic actuator816is connected to the printing tray assembly822with a connection piece818that moves the entire printing tray assembly822along on a track826on the printing platform814. The z-axis hydraulic actuator820is connected to the printing tray assembly822and moves along with it while moving the printing tray824along a track828on the printing tray assembly822. This precisely positions the printing tray824for three-dimensional printing.

FIG. 15Dis a top view of the hydraulically driven three-dimensional printing device800where the printing tray824is moved along the x-axis and z-axis with corresponding hydraulically driven actuators816′ and820′. In this embodiment the x-axis hydraulic actuator816′ has extended and moved the printing tray assembly822horizontally across the printing platform814. The z-axis hydraulic actuator820′ has extended and moved the printing tray824′ in the z-axis direction across the printing tray assembly822. This precisely positions the printing tray for three-dimensional printing.

FIGS. 16A-16Gshow several embodiments of printing with multiple degrees of freedom.

FIG. 16Ais a side view of a three-dimensional printer840with a hydraulically- and/or pneumatically-linear-actuator-driven printing platform846. In this embodiment at least two hydraulically- and/or pneumatically-driven linear actuators842and844are positioned on either side of the printing platform846. The hydraulically- and/or pneumatically-driven linear actuators842and844are uniformly extended and raise the printing platform846up, thereby positioning the printing tray848in the precise location for printing via the printing head850.

FIG. 16Bis a side view of a three-dimensional printer840′ with a hydraulically- and/or pneumatically-linear-actuator-driven printing platform846′. The hydraulically- and/or pneumatically-driven linear actuators842′ and844′ are extended at different heights resulting in the printing platform846′ and printing tray848′ to be positioned at an angle in relation to the printing head850. The positioning of the printing tray848′ at a plurality of angles allows for additional degrees of freedom and increased flexibility for printing along multiple axes for three-dimensional printing with respect to a standard three-axis-coordinate printing system. This will assist in the building of structures, such as bioprinted structures, which may require additional degrees of freedom for proper formation of a biological manufactured product.

FIG. 16Cis an alternate embodiment of a side view for a three-dimensional dimensional printer840″ with a hydraulically- and/or pneumatically-linear-actuator-driven printing platform846″. The hydraulically- and/or pneumatically-driven linear actuators842″ and844″ are extended into different positions at different heights resulting in the printing platform846″ and printing tray848″ to be positioned at an alternate angle in relation to the printing head850with respect to the angle shown inFIG. 16B.

FIG. 16Dis a side view of a three-dimensional dimensional printer860with a printing head866featuring an articulating axis joint868. The articulating axis joint868allows for the positioning of the printing head dispenser870at a plurality of angles in relation to the printing tray864located on the printing platform862, which allows for additional degrees of freedom and increased flexibility for printing along multiple axes for three-dimensional printing with respect to a standard three-axis-coordinate printing system. The movement of the articulating axis joint868on the printing head dispenser870may be driven by a hydraulic, pneumatic, magnetic, electric, and/or other internal/external force. The articulating axis joint868may be a hinge joint or a ball and socket joint.

FIG. 16Eis an alternate embodiment (with respect to view D′) of a side view for a three-dimensional dimensional printer860′ with a printing head866featuring an articulating axis joint868′. The articulating axis joint868′ is moved into an alternate position which allows for the printing head dispenser870′ to be positioned at a different angle in relation to the printing tray864.

FIG. 16Fis a side view of a three-dimensional dimensional printer872with a printing head874featuring a plurality of articulating axis joints876and878. The at least two articulating axis joints876and878on the printing head874allow for the positioning of the printing head dispenser880at a plurality of angles in relation to the printing tray888. A three-dimensional printer with a plurality of articulating axis joints876and878allows for the top portion of the printing head874to remain in a fixed position through a fixed port on the top of the single-use container as well as for a fixed position of the printing tray888, while allowing the multi-axis positioning of the printing head without the required movements and positioning by a gantry assembly.

FIG. 16Gis a side view of a three-dimensional dimensional printer872with a hydraulically- and/or pneumatically-linear-actuator-driven printing platform886and a printing head874featuring a plurality of articulating axis joints876and878. The at least two articulating axis joints876and878on the printing head874allow for the positioning of the printing head dispenser880at a plurality of angles in relation to the printing tray888. The hydraulically- and/or pneumatically-driven linear actuators882and884are extended at different heights resulting in the printing platform886and printing tray888to be positioned at an angle in relation to the printing head dispenser880. The multi-axis positioning of the printing tray888at a plurality of angles as well as the multi-axis positioning of the printing head dispenser880allows for additional degrees of freedom and increased flexibility for printing with respect to a standard three-axis-coordinate printing system.

FIGS. 17A-17Gshow several embodiments of moving print platforms for printing with multiple degrees of freedom.

FIG. 17Ais a top view of a printer chamber setup900containing a printing tray902with a rotational shaft at the center and a hydraulic, pneumatic, electric, and/or magnetically controlled linear actuator904set at an angle with the printing tray902, where the linear actuator shaft connects to the printing tray902with a rotating connection joint906. The linear actuator shaft is in an idle position.

FIG. 17Bis a top view of a printer chamber setup900′ where the linear actuator904′ shaft is extended, thereby pushing on the rotating connection joint906′ and causing the printing tray902′ to rotate along the shaft at the center away from the linear actuator904′. This extension of the linear actuator904′ causing the rotation of the printing tray902′ allows for an additional degree of freedom for movement during the printing process from a printing head. This rotational movement may allow a fixed printing head previously confined to a standard three-axis control setup by gantry positioning to now provide the ability to make curved features, which are present throughout biological and/or biologically-inspired components but are difficult to produce at high resolution with a standard three-axis positioning system.

FIG. 17Cis an alternate view of a printer chamber setup900″ where the linear actuator904″ shaft is contracted, thereby pulling on the rotating connection joint906″ and causing the printing tray902″ to rotate along the shaft at the center towards the linear actuator904″. This contraction of the linear actuator904″ allows for a rotation of the printing tray902″ during the printing process.

FIG. 17Dis a top view of a printer chamber setup908where the square printing tray910undergoes a rotation movement912via a moving shaft at the center of the square printing tray910. The center shaft of the square printing tray910may be powered by a rotational motor driven by hydraulic, pneumatic, electric, or magnetically controlled methods (not shown). This rotational movement may allow a fixed printing head to make curved, circular, elliptical, parabolic, hyperbolic, or spheroidal features.

FIG. 17Eis a top view of a printer chamber setup914where the circular printing tray916undergoes a rotation movement918via a moving shaft at the center of the circular printing tray916. The center shaft of the circular printing tray916may be powered by a rotational motor driven by hydraulic, pneumatic, electric, or magnetically controlled methods (not shown).

FIG. 17Fis a side view of a printer chamber setup920where the printing tray924is connected to a rotating and articulating joint926on the printing platform922. The rotating and articulating joint926may be a hinge joint or a ball and socket joint. The rotating and articulating joint926can move in a rotational direction928and/or a vertical motion930, which changes the position of the printing tray924to a plurality of angles relative to a fixed printing head. The rotating and articulating joint926may be powered by a rotational motor driven by hydraulic, pneumatic, electric, or magnetically controlled methods (not shown).

FIG. 17Gis a side view of a printer chamber setup932where the printing tray934is connected to a rotating and articulating joint936on the printing platform. The rotating and articulating joint936can move in a rotational direction and/or a vertical motion942, which is controlled by a plurality of linear actuators938and940connected to the underside of the printing tray934. The linear actuators938and940may be extended or contracted to different heights, causing the position of the printing tray934to be raised or lowered at a plurality of angles relative to a fixed printing head. The connection of the linear actuators938and940to the underside of the printing tray934may be implemented via a track (not shown), which allows the linear actuator shafts to move freely when the printing tray934is rotated while remaining connected, in order to cause the movement of the printing tray934to the desired angle during the printing process. The rotating and articulating joint936as well as the linear actuators938and940may be powered by a rotational motor driven by hydraulic, pneumatic, electric, or magnetically controlled methods (not shown).

FIGS. 18A-18Mshow several embodiments of printer designs. The embodiments of the printer designs are simplified to show the variety of methods for dispensing material within a sterile chamber. More complex designs of each of these embodiments may contain sterilizing grade vent filters, sterile drains, transfer hatches, and other accessories described in the other figures.

FIG. 18Ais a side view of a printer chamber setup1000containing a three-axis gantry positioning system utilizing at least one vertical linear actuator1002to adjust the height of the printing platform1004. The printing platform1004may support a horizontal linear actuator1006, a second horizontal linear actuator1008orthogonal to linear actuator1006and a moving printing tray1010. The horizontal linear actuator1006may be utilized to position the printing tray1010along the x-axis and the diagonal linear actuator1008may be utilized to position the printing tray1010along the z-axis. The three-axis gantry positioning system is utilized for printing with a fixed printing head1012within the sterile printer chamber setup1000. The linear actuators may be driven by hydraulic, pneumatic, electric, or magnetically controlled methods (not shown). This embodiment is a minimal design where only three linear actuators may be utilized to position the 3-axis positioning system, where in other embodiments a plurality of actuators including more than three actuators may be utilized.

FIG. 18Bis a side view of a printer chamber setup1020containing a hybrid three-axis gantry positioning system utilizing at least one vertical inflatable bag assembly1022to adjust the height of the printing platform1024. The vertical inflatable bag assembly1022may contain a plurality of bags, which may be filled with a fluid (air, gas, or liquid) to vertically position the printing platform1024. The printing platform1024may support a horizontal actuator1026, a second horizontal actuator1028orthogonal to horizontal actuator1026and a moving printing tray1030. The horizontal actuator1026may be utilized to position the printing tray1030along the x-axis and the second horizontal actuator1028may be utilized to position the printing tray1030along the z-axis. The three-axis gantry positioning system is utilized for printing with a fixed printing head1032within the sterile printer chamber setup1020. The inflatable bag assembly1022may be driven by hydraulic, pneumatic, or chemical methods (not shown), while the actuators may be driven by hydraulic, pneumatic, electric, or magnetically controlled methods (not shown). In alternate embodiments a plurality of bag assemblies and actuators may be utilized.

FIG. 18Cis a side view of a printer chamber setup1040containing a hybrid three-axis gantry positioning system utilizing at least one vertical magnetically-controlled assembly1042to adjust the height of the printing platform1044. The magnetically-controlled assembly1042utilizes magnets, ferrimagnets, rare earth magnets, magnetic fields, or ferrofluids on the exterior of the printer chamber setup1040, which are paired with an equivalent magnet on the printing platform1044within the interior of the printer chamber setup1040. The printing platform1044may support a horizontal actuator1046, a second horizontal actuator1048orthogonal to horizontal actuator1046and a moving printing tray1050. The horizontal actuator1046may be utilized to position the printing tray1050along the x-axis and the second horizontal actuator1048may be utilized to position the printing tray1050along the z-axis. The three-axis gantry positioning system is utilized for printing with a fixed printing head1052within the sterile printer chamber setup1040. The magnetically controlled assembly1042may be driven by mechanical, electric methods (not shown) or through the generation of magnetic fields (not shown), while the actuators may be driven by hydraulic, pneumatic, electric, or magnetically controlled methods (not shown). In alternate embodiments a plurality of magnetically-controlled assemblies and actuators may be utilized.

FIG. 18Dis a side view of a printer chamber setup1060containing a multi-axis positioning system utilizing a robotic arm1062for positioning a printing tray1074with a plurality of articulating axis joints internal to the printer chamber setup1060. In one embodiment the robotic arm1062may be controlled by utilizing an actuator1064to rotate a rotating base1066. An additional actuator1068pushes on a support connected to an articulating joint moving the position of a printing platform1072and the printing tray1074vertically. An additional actuator1070pushes on a support connected to an articulating joint moving the position of the printing platform1072and printing tray1074at an angle in relation to the fixed printing head1076. The actuators may be driven by hydraulic, pneumatic, electric, or magnetically controlled methods (not shown). In other embodiments a plurality of additional actuators may be used to position the printing tray1074on the printing platform1072.

FIG. 18Eis a side view of a printer chamber setup1080containing a hybrid three-axis gantry positioning system utilizing at least one vertical platform jack assembly1082to adjust the height of a printing platform1090. In this embodiment the platform jack assembly1082contains a platform jack assembly screw1083in a fixed vertical position (known as a low-profile or offset platform jack) to control the positioning of the platform jack scissoring and the printing platform1090height. The platform jack assembly screw1083is connected to a connection component on the interior of the printer chamber setup1080via an extension assembly1084which may be a scissoring assembly, a suspension cylinder assembly, a hydraulic/pneumatic cylinder assembly, or a spring assembly. As the platform jack assembly1082is raised or lowered, the platform jack assembly screw1083changes horizontal position, thereby forcing the extension assembly1084inside of the sealed printer chamber1080. The connection component assembly1086contains the interior connection component that drives the platform jack assembly screw1083, the exterior connection component that connects to an external motor1088to turn the interior connection component, and a chamber interface component that connects the exterior of the chamber with the interior. The exterior connection component of the connection component assembly1086connects to the exterior of the printer chamber setup1080through magnets, or an internal and/or external shaft. In the magnetically-driven embodiment the chamber interface component may comprise a rigid plastic material, seals, and bearings to prevent wear along the interface between the exterior and interior of the chamber. In the internal and/or external shaft embodiment an external shaft may be inserted into the interior connection component (or vice versa) through the chamber interface component, which may comprise a rigid plastic material, seals, and bearings to prevent wear along the interface between the exterior and interior of the chamber. An external motor1088contains an external magnetic assembly (not shown) or an external shaft assembly (not shown), which is inserted into the connection component assembly1086to drive the platform jack assembly screw1083in order to change the position of the printing platform1090. The printing platform1090may support a horizontal actuator1092, a second horizontal actuator1094orthogonal to horizontal actuator1092and a moving printing tray1096. The horizontal actuator1092may be utilized to position the printing tray1096along the x-axis and the second horizontal actuator1094may be utilized to position the printing tray1096along the z-axis. The three-axis gantry positioning system is utilized for printing with a fixed printing head1098within the sterile printer chamber setup1080. The actuators may be driven by hydraulic, pneumatic, electric, or magnetically controlled methods (not shown). Various embodiments of the platform jack positioning may be utilized to control different axis positioning of the printer assembly.

FIG. 18Fis a side view of a printer chamber setup1100containing a hybrid three-axis gantry positioning system utilizing at least one vertical platform jack assembly1102to adjust the height of a printing platform1110. In this embodiment the platform jack assembly1102contains a standard platform jack assembly with a platform jack assembly screw1103that moves in both vertical and horizontal directions during the rotation of the screw1103, which changes the height of the jack scissoring and of the printing platform1110. The platform jack assembly screw1103is connected to a connection component on the interior of the printer chamber setup1100via a jointed extension assembly1104, which may be a jointed scissoring assembly, a jointed suspension cylinder assembly, a jointed hydraulic/pneumatic cylinder, or a jointed spring assembly. The jointed extension assembly1104comprises an extension element connected to a hinged or ball and socket joint at either end, or a hinged or ball and socket joint in the middle of the assembly. As the platform jack assembly1082is raised or lowered, the platform jack assembly screw1103changes position vertically and horizontally, forcing the extension assembly1104inside of the sealed printer chamber1100. The jointed extension assembly1104allows for the rotational force to act on the platform jack assembly screw1103even when the screw1103changes position horizontally or vertically. The connection component assembly1106contains the interior connection component that drives the platform jack assembly screw1103, the exterior connection component that connects to an external motor1108to turn the interior connection component, and a chamber interface component that connects the exterior of the chamber with the interior. The exterior connection component of the connection component assembly1106connects to the exterior of the printer chamber setup1100through magnets, or an internal and/or external shaft. In the magnetically-driven embodiment the chamber interface component may comprise a rigid plastic material, seals, and bearings to prevent wear along the interface between the exterior and interior of the chamber. In the internal and/or external shaft embodiment an external shaft may be inserted into the interior connection component through the chamber interface component, which may consist of a rigid plastic material, seals, and bearings to prevent wear along the interface between the exterior and interior of the chamber. An external motor1108contains an external magnetic assembly (not shown) or an external shaft that is inserted into the connection component assembly1106to drive the platform jack assembly screw1103to change the position of the printing platform1110. The printing platform1110may support a horizontal actuator1112, a second horizontal actuator1114orthogonal to horizontal actuator1112and a moving printing tray1116. The horizontal actuator1112may be utilized to position the printing tray1116along the x-axis and the second horizontal actuator1114may be utilized to position the printing tray1116along the z-axis. The three-axis gantry positioning system is utilized for printing with a fixed printing head1118within the sterile printer chamber setup1100. The actuators may be driven by hydraulic, pneumatic, electric, or magnetically controlled methods (not shown). Various embodiments of the platform jack positioning may be utilized to control different axis positioning of the printer assembly.

FIG. 18Gis a side view of a printer chamber setup1120containing a hybrid three-axis gantry positioning system utilizing at least one vertical platform jack assembly1122to adjust the height of a printing platform1136. In this embodiment the platform jack assembly1122contains a standard platform jack assembly with a platform jack assembly screw1123that moves in both vertical and horizontal directions during the rotation of the screw, which changes the height of the jack scissoring and of the printing platform1136. The platform jack assembly screw1123is connected to a connection component on the interior of the printer chamber setup1120via an extension assembly1124, which may be a scissoring assembly, a suspension cylinder assembly, a hydraulic/pneumatic cylinder, or a spring assembly. As the platform jack assembly1122is raised or lowered, the platform jack assembly screw1123changes position horizontally, forcing the extension assembly1124inside of the sealed printer chamber1100. The platform jack assembly screw1123additionally changes position vertically, causing the vertical movement of connection component assembly. The connection component assembly1126in this embodiment utilizes a separate exterior platform jack1132with a platform jack assembly screw in a fixed vertical position (known as a low-profile or offset platform jack) where an external motor1130is raised with the external platform jack1132to match the height of the internal platform jack1122. The external platform jack1132itself contains its own platform jack motor1134for adjusting the height of the external motor1130. The height adjustment from both external motors1130and1134may be computer-controlled to keep the internal platform jack1122and the external platform jack1132properly aligned. Alternatively or additionally a detection system such as a camera detection system with a target may be utilized to keep the assembly properly aligned. The exterior connection component1128of the connection component assembly connects to the exterior of the printer chamber setup1120through magnets, or an internal and/or external shaft. In the magnetically-driven embodiment the chamber interface component1126may comprise a rigid plastic material, seals, and bearings to prevent wear along the interface between the exterior and interior of the chamber. In the internal and/or external shaft embodiment an external shaft may be inserted into the interior connection component through the chamber interface component1126, which may comprise a rigid plastic material, seals, and bearings to prevent wear along the interface between the exterior and interior of the chamber. The external motor1130contains an external magnetic assembly (not shown) or an external shaft that is inserted into the connection component assembly to drive the platform jack assembly screw1123to change the position of the printing platform1136. The printing platform1136may support a horizontal actuator1138, a second horizontal actuator1140orthogonal to horizontal actuator1138and a moving printing tray1142. The horizontal actuator1138may be utilized to position the printing tray1142along the x-axis and the second horizontal actuator1140may be utilized to position the printing tray1142along the z-axis. The three-axis gantry positioning system is utilized for printing with a fixed printing head1144within the sterile printer chamber setup1120. The actuators may be driven by hydraulic, pneumatic, electric, or magnetically controlled methods (not shown). Various embodiments of the platform jack positioning may be utilized to control different axis positioning of the printer assembly.

FIG. 18His a side view of a printer chamber setup1150containing a hybrid three-axis gantry positioning system utilizing at least one vertical linear actuator1152to adjust the height of a printing platform1154. The printing platform1154may support a printing tray1156which may move along a series of tracks for positioning. In this embodiment a plurality of soft robotic actuators1158and1160, which cause the soft robotics material to curl when pressurized with hydraulic or pneumatic fluid pressure, are utilized to precisely move the printing tray1156in relation to the fixed printing head1164. The soft robotic actuators1158and1160may be positioned to push and pull the printing tray1156along a series of tracks (not shown) on the printing platform1154in the two horizontal positions. In this embodiment the soft robotic actuator1158is in a resting position, while the soft robotic actuator1160is in the deployed position when fluid pressure enters into the fluid assembly1162. The use of soft robotic actuators is advantageous in that there are no rigid materials if a non-rigid chamber is utilized, they are not made from complex components, and they are readily sterilizable. The three-axis gantry positioning system is utilized for printing with a fixed printing head1164within the sterile printer chamber setup1150. The linear actuators may be driven by hydraulic, pneumatic, electric, or magnetically controlled methods (not shown). In alternate embodiments a plurality of soft robotic actuators and linear actuators may be utilized.

FIG. 18Iis a side view of a printer chamber setup1170containing a hybrid multiaxis positioning system utilizing at least one vertical linear actuator1172to adjust the height of a printing platform1174. The printing platform1174may support a printing tray1176, which may be in a fixed position or move along a series of tracks for positioning. In this embodiment a plurality of soft robotic actuators1178and1180, which cause the soft robotic material to curl when pressurized with hydraulic or pneumatic fluid pressure, are utilized to precisely move a plurality of printing heads1184with a fixed position at the top of the chamber into position above the printing tray1176for dispensing material. The plurality of soft robotic actuators1178and1180may be positioned in a plurality of directions above the printing tray1176for precisely dispensing material. In this embodiment the soft robotic actuator1178is in an actuated position and moves a printing head into a precise position above the printing tray1176. The soft robotic actuator1180is in an actuated position and moves a different printing head into another precise position above the printing tray1176. The soft robotic actuators1178and1180are in the deployed position when fluid pressure enters into the fluid assembly1182. The linear actuators may be driven by hydraulic, pneumatic, electric, or magnetically controlled methods (not shown). In alternate embodiments a plurality of soft robotic actuators, printing heads, and linear actuators may be utilized.

FIG. 18Jis a side view of a printer chamber setup1190containing a hybrid multiaxis positioning system utilizing at least two vertical soft robotic actuators1194and1196to adjust the height of a printing platform1192. The printing platform1192may support a horizontal soft robotic bellows actuator1202, a second horizontal soft robotic bellows actuator (not shown) and a moving printing tray1204. In this embodiment a plurality of soft robotic actuators1194and1196, which cause the soft robotic material to curl when pressurized with hydraulic or pneumatic fluid pressure, are utilized to precisely move the printing platform1192in a vertical direction. In this embodiment the soft robotic actuators1194and1196are in an actuated position and move the printing platform1192into a precise position below the fixed printing head1206. The soft robotic actuators1194and1196are in the deployed position when fluid pressure enters into the fluid assemblies1198and1200, respectively. The horizontal soft robotic bellows actuator1202is a soft robotic linear actuator that expands the bellows when actuated under fluid pressure. The soft robotic bellows actuator1202may be utilized to position the printing tray1204along the x-axis. An additional horizontal linear actuator (not shown) may be utilized to position the printing tray1204along the z-axis. Alternatively the soft robotic actuators controlling the movement of the printing platform1192along the y-axis may additionally be utilized to position the printing platform1192along an additional axis such as the x-axis or z-axis. The soft robotic actuators may be driven by hydraulic and/or pneumatic fluid pressure.

FIG. 18Kis a side view of a printer chamber setup1210containing a hybrid multiaxis positioning system utilizing a fixed printing platform1212and a tethered multiaxis printing device1216. In this embodiment the tethered multiaxis printing device1216comprises a plurality of soft robotic appendages1218, which are attached to a core of the device that contains the printing head dispenser1220and the printing head assembly1222. The printing head assembly1222is tethered to the top of the printer chamber setup1210through a port connection1226that is utilized to feed material to the tethered multiaxis printing device1216. The printing platform1212may support a printing tray1214, which may be in a fixed position or move along a series of tracks for positioning. The tethered multiaxis printing device1216utilizes a plurality of soft robotic appendages1218, where the soft robotic material curls when pressurized with hydraulic or pneumatic fluid pressure, to precisely position the printing head dispenser1220above the printing tray1214. The plurality of soft robotic appendages1218may also be utilized to push and pull the printing tray1214along a series of tracks (not shown) on the printing platform1212in the two horizontal directions. In this embodiment a plurality of soft robotic appendages1218moves when fluid pressure enters into the fluid assemblies1224. The soft robotic appendages may be driven by hydraulic and/or pneumatic fluid pressure.

FIG. 18Lis a side view of a printer chamber setup1230containing a multiaxis positioning system utilizing a fixed printing platform1232and an untethered multiaxis printing device1236. In this embodiment the untethered multiaxis printing device1236comprises a plurality of soft, rigid, and/or rigid coated with soft material robotic appendages1238, which are attached to a core of the device that contains the printing head dispenser1240and the printing head assembly1242, which contains a storage reservoir to hold the materials to dispense. A plurality of storage reservoirs may be utilized for dispensing solid, liquid, gas or mixed materials, where compressed gas or air may be used to drive and regulate the dispensing of solid or liquid materials including cells in a liquid suspension. The printing head assembly1242may contain a heater for heating the material prior to dispensing. The printing platform1232may support a printing tray1234, which may be in a fixed position or move along a series of tracks for positioning. The untethered multiaxis printing device1236utilizes a plurality of robotic appendages1238, where the appendages move in response to the movement of the drive mechanism, which may be driven by hydraulic, pneumatic, electric, or magnetically controlled methods (not shown). The drive mechanism may contain its own reservoir within the untethered multiaxis printing device1236, including a reservoir for hydraulic and/or pneumatic fluid pressure, an internal pump for generating such pressure, a battery pack, a plurality of magnets or other elements. The battery may be charged by inductive charging, directed charging through microwave, laser, or other line-of-sight radiation, solar cell or other wireless charging methods. The untethered multiaxis printing device1236may additionally move to a location inside the interior of the printer chamber setup1230, where it can refill the internal reservoirs for solid, liquid, gas, or mixed inputs as well as recharge an internal reservoir for the drive mechanism, including charging a pressurized hydraulic and/or pneumatic reservoir, a battery through direct charging with energized contacts, inductive charging, or other mechanical energy storage mechanism (winding). The untethered multiaxis printing device1236plurality of robotic appendages1238may be utilized to position the at least one printing head dispenser1240above the printing tray1234. The untethered multiaxis printing device1236may contain a plurality of printing heads, each of which has a different function and dispenses material from a plurality of corresponding internal reservoirs. The plurality of robotic appendages1238may also be utilized to push and pull the printing tray1234along a series of tracks (not shown) on the printing platform1232in the two horizontal directions. The untethered multiaxis printing device1236has the advantage of being completely internal to the printer chamber setup1230without any external connections or tethering. Additionally the setup is advantageous as the untethered multiaxis printing device1236can provide multiaxis printing for a plurality of printing trays by moving to each one independently.

FIG. 18Mis a side view of a printer chamber setup1250containing a plurality of multiaxis positioning devices1256,1258,1260, and1262utilizing a fixed printing platform1252and a printing tray1254. In this embodiment the plurality of untethered multiaxis printing devices1256,1258,1260, and1262operate as described in View ‘L’ with the exception of the potential to be individualized and capable of performing different functions. A plurality of storage reservoirs (not shown) within the plurality of multiaxis positioning devices1256,1258,1260, and1262may be utilized for dispensing solid, liquid, gas or mixed materials, where compressed gas or air may be used to drive and regulate the dispensing of solid or liquid materials including cells in a liquid suspension. In this embodiment a single untethered multiaxis printing device1256,1258,1260, or1262may contain a single reservoir and a single material to dispense out of the printing head dispenser. The material dispensed from each untethered multiaxis printing device1256,1258,1260, or1262may be different for each device. The printing platform1252may support the printing tray1254, which may be in a fixed position or move along a series of tracks for positioning. In this embodiment the untethered multiaxis printing device1262contains robotic appendages with magnetic, conductive, or ferrous material at the base that comes in contact with the surface for locomotion. Magnetic plates1264, ferrous material, and/or magnetic fields may be placed directly external to the printer chamber setup1250, where the multiaxis positioning devices1256,1258,1260, and1262may utilize the full surface area of the interior of the chamber for locomotion, positioning, charging, refilling reservoirs, or other operational functions. The untethered multiaxis printing devices1256,1258,1260, and1262may be sterilized within the printer chamber setup1250as a complete assembly or may be sterilized using an alternate process (such as chemical sterilization) and aseptically inserted into the interior of the printer chamber setup1250. The untethered multiaxis printing devices1256,1258,1260, and1262have the advantage of being simpler and dedicated to the material inputs that they hold, they can be redundant in case of operating failure inside of sterile chamber, they can self-position and print (dispense materials) over a plurality of printing trays, and they can be replaceable in that additional devices may be aseptically inserted into the interior of the printer chamber setup1250.

FIGS. 19A-19Fshow an alternate embodiment of the connections steps to insert the printing head into the sterile three-dimensional assembly body.

FIG. 19Ais a side view of an embodiment that shows a sterilized three-dimensional printer assembly body2000, with a rigid or flexible wall, containing a printing head insertion assembly2002. The portion of the printing head insertion assembly2002internal to three-dimensional printer assembly body2000contains an interface assembly2006in the closed position and an internal dip tube2004. The external portion of the printing head insertion assembly2002contains a removable cap2012, a fluid inlet port2008, and a fluid outlet port2010.

FIG. 19Bis a side view of the removable cap2012′ being removed from the printing head insertion assembly2002and the electronically-controlled printing head2014being inserted into the assembly.

FIG. 19Cis a side view of the electronically-controlled printing head2014inserted into the printing head insertion assembly2002and twisted into place2016in a first position.

FIG. 19Dis a side view of the printing head insertion assembly2002with a tubing piece2018inserted into the fluid inlet port2008, wherein a chemical sterilant or sanitizer2020fills the printing head insertion assembly2002and the interface assembly2006, thereby sterilizing the internal section of the electronically-controlled printing head2014. The chemical sterilant or sanitizer2020can comprise a liquid like 30% hydrogen peroxide, or may comprise a gas sterilant such as vaporized hydrogen peroxide or ethylene oxide.

FIG. 19Eis a side view of the printing head insertion assembly2002with a tubing piece2022inserted into the fluid outlet port2010, wherein the chemical sterilant or sanitizer is removed by vacuum pressure after the period of time required to sterilize the printing head2014is completed. The internal dip tube2004can be utilized to remove a fluid chemical sterilant or sanitizer or condensate from a gas chemical sterilant.

FIG. 19Fis a side view of the printing head insertion assembly where the electronically-controlled printing head2014is twisted2024into a second position, where either the printing head2014is screwed down and breaks through the interface assembly2006′ into the interior of the three-dimensional printer assembly body or the interface assembly2006′ opens up via a mechanical mechanism (not shown) in multiple directions (like a flower petal or gates). In this way, the sterilized electronically-controlled printing head2014is allowed access to the interior of the three-dimensional printer assembly body2000. The sterilized electronically-controlled printing head2014is ready to print within the three-dimensional printed assembly body2000.

FIGS. 20A-20Eshow another embodiment of the connections steps to insert the printing head into the sterile three-dimensional assembly body.

FIG. 20Ais a side view of an embodiment that shows a sterilized three-dimensional printer assembly body2030, with a rigid or flexible wall, containing a printing head insertion assembly2032. The printing head insertion assembly2032may comprise an aseptic connection strip2034, which may contain a sterilizing grade filter membrane and be held in place within the assembly with a seal, an aseptic connection strip guide2036to guide the aseptic connection strip2034as it is removed, and excess material2038from the aseptic connection strip2034for the operator to pull after the connection has taken place. The external portion2040of the printing head insertion assembly2032contains an aseptic connector with a removal strip2042, which may additionally contain a sterilizing grade filter and be held in place within the assembly

FIG. 20Bis a side view of the sterilized three-dimensional printer assembly body2030containing a printing head insertion assembly2032. A sterilized printing head assembly2044, which itself may be rigid or flexible, contains an aseptic connector2046, which may additionally contain a sterilizing grade filter and be held in place within the assembly2044with a seal, an internal sterilized printing head2048, and a lever assembly2050to pull the internal sterilized printing head2048into the three-dimensional printer assembly body2030. The sterilized printing head2048may be single-use or multi-use and capable of being re-sterilized.

FIG. 20Cis a side view of the sterilized printing head2044where the body is twisted into place locking the at least two aseptic connectors2040and2046into place. The operator pulls the aseptic connection strips2042and2052, which may contain sterilizing grade filter membranes, in direction2056. The removal of the two aseptic connection strips2042and2052opens up the sterilized printing head2044into the printing head insertion assembly2032. The aseptic connection strip2034, which may contain a sterilizing grade filter membrane, may remain in place if the aseptic connection between aseptic connectors2040and2046was not properly connected or the sterilized printing head2044integrity was compromised. The integrity of this assembly may be integrity tested with an integrity testing device (not shown) through a vented connection with a sterilizing grade filter (not shown) prior to proceeding with the next steps of removing the aseptic seal within the sterilized three-dimensional printer assembly body2030.

FIG. 20Dis a side view of the sterilized printing head assembly2044where the lever assembly2050is depressed and pulls the internal sterilized printing head2048into the printing head insertion assembly2032. The printer material feed tubing assembly2058, which brings material inputs (such as fluids with biologically active products or structural material) to the sterilized printing head2048, may be coiled or stored within the sterilized printing head assembly2044and sterilized along with the assembly2044. A port2060in the sterilized printing head assembly2044serves as an interface between the feed tubing in the interior and exterior of the setup. Alternatively the tubing line to bring the input material to the sterilized printing head2048may be aseptically connected to the sterile assembly. The operator pulls the excess material2038from the aseptic connection strip2034away from the printing head insertion assembly2032. The aseptic connection strip2034, which may contain a sterilizing grade filter membrane, is pulled through the aseptic connection strip guide2036.

FIG. 20Eis a side view of the printing head insertion assembly2032where the removal of this aseptic connection strip2034opens up the sterilized printing head2048to the internal sterilized chamber of the three-dimensional printer assembly body2030. The sterilized printing head2048may now print to the internal printing tray within the unit, where additional inputs are brought through internal tubing2058.

FIGS. 21A-21Eshows an alternate embodiment of the connections steps to insert a sterilized printing head with a bellows assembly into the sterile three-dimensional assembly body.

FIG. 21Ais a side view of an embodiment that shows a sterilized three-dimensional printer assembly body2068, with a rigid or flexible wall, containing a printing head insertion assembly2070comprising an aseptic connection strip2072held in place within the assembly with a seal, an aseptic connection strip guide2074to guide the aseptic connection strip2072as it is removed, and excess material2076from the aseptic connection strip2072for the operator to pull after the connection has taken place. A sterilized printing head assembly with a bellows assembly2078, which itself may be rigid or flexible, contains an aseptic connector, which may additionally contain a sterilizing grade filter and be held in place within the assembly with a seal, and an internal sterilized printing head2082. The sterilized printing head assembly with a bellows assembly2078uses the bellows assembly to push the sterilized printing head2082into the printing head insertion assembly2070within the three-dimensional printer assembly body2068. The sterilized printing head2082may be single-use or multi-use and capable of being re-sterilized.

FIG. 21Bis a side view of the sterilized printing head assembly with a bellows assembly2078where the body is twisted2084into place locking the at least two aseptic connectors into place. The operator pulls the aseptic connection strips2086and2088, which may contain sterilizing grade filter membranes, in direction2090. The removal of the two aseptic connection strips2086and2088opens up the sterilized printing head assembly with a bellows assembly2078into the printing head insertion assembly2070. The aseptic connection strip2072, which may contain a sterilizing grade filter membrane, may remain in place if the aseptic connection between aseptic connectors was not properly connected or the sterilized printing head2082integrity was compromised. The integrity of this assembly may be integrity tested with an integrity testing device (not shown) through a vented connection with a sterilizing grade filter (not shown) prior to proceeding with the next steps of removing the aseptic seal within the sterilized three-dimensional printer assembly body2068.

FIG. 21Cis a side view of the sterilized printing head assembly with a bellows assembly2078where the operator pushes on the sterilized printing head assembly with a bellows assembly2078in direction2092compressing the bellows assembly and pushing the sterilized printing head2082into the printing head insertion assembly2070.

FIG. 21Dis a side view of the sterilized printing head assembly with a bellows assembly2078where the assembly is twisted in direction2094and locked into place. The operator pulls in direction2096the excess material2076from the aseptic connection strip2072away from the printing head insertion assembly2070. The aseptic connection strip2072is pulled through the aseptic connection strip guide2074.

FIG. 21Eis a side view of the printing head insertion assembly2070where the removal of this aseptic connection strip2072opens up the sterilized printing head2082to the internal sterilized chamber of the three-dimensional printer assembly body2068. The sterilized printing head2082may now print to the internal printing tray.

FIG. 22shows an embodiment of a manually-controlled robotic arm within a sterilized three-dimensional assembly chamber. The chamber and robotic arm control assembly3000comprises a sterilized chamber3002that contains at least one robotic arm assembly3004, a port3017to connect a control assembly3030with the robotic arm assembly3004, and a transfer hatch3040for the chamber3002. In this embodiment the robotic arm assembly3004utilizes a plurality of actuators to control the positioning and movements of the assembly. The robotic arm assembly3004, including the plurality of actuators, may be made out of sterilizable (gamma irradiatable and/or autoclavable) plastic materials and rubberized seals. In this embodiment the robotic arm assembly3004may be controlled by utilizing an actuator3008to rotate a rotating base3006. An additional actuator3010pushes on a support connected to an articulating joint moving the position of the arm. An additional actuator3012pushes on a support connected to an articulating joint moving the position of the robotic grippers3016. An additional actuator3014pushes on a support connected to an articulating joint that opens and closes the robotic grippers3016. The actuators may be driven by hydraulic, pneumatic, electric, or magnetically controlled methods (not shown). In this embodiment the robotic arm control assembly3030is a manual controller external to the sterilized chamber3002and utilizes hydraulic and/or pneumatic fluid pressure to control the movements of the robotic arm assembly3004internal to the sterilized chamber3002. The manual robotic arm control assembly3030utilizes a plurality of hydraulic and/or pneumatic tubing lines3018to move fluid through the port3017into the actuators located on the robotic arm assembly3004to control the movements. The tubing lines3018are charged with sterile hydraulic and/or pneumatic fluid pressure through the filling assembly3020. In this embodiment the tubing lines are filled with a hydraulic fluid, which is for example sterile filtered water. The sterile filtered water enters into the chamber and robotic arm control assembly3000after sterilization and is setup for use by an operator. Purified water enters through the tubing line3022and passes through a sterilizing grade filter3024into the filling assembly3020, which may serve as a manifold to completely fill each of the individual tubing lines3018. The tubing lines3018may be cleared of air through a sterilizing grade vent filter3026, which allows the displaced air to vent to the atmosphere as it is displaced by sterile filtered water entering into the assembly. Additional sterile filtered water, other fluids, or, in other embodiments, pneumatic air or gas pressure may be added to the tubing lines3018via the filling assembly3020in case of leakage or loss of pressure. In this basic embodiment a manual robotic arm control assembly3030is utilized to control the movements of the robotic arm assembly3004. A plurality of hydraulic and/or pneumatic pistons3034are arranged in a piston assembly3032that comprises cylinders filled with fluid, and the piston heads3038are pushed and/or pulled by the operator. The movement of one of the piston heads3038affects the movement of a seal3036internal to the piston and causes the displacement of the internal fluid through the tubing lines3028and3018, resulting in the movement of the robotic arm assembly3004. The purpose of the robotic arm assembly3004is to manipulate and move objects internal to the sterilized chamber3002. This has the benefits of moving, storing, stacking or manipulating the objects inside of the sterilized chamber3002in ways not possible before. The transfer hatch3040may be utilized to remove objects aseptically from the sterilized chamber3002and/or to connect to additional sterilized chambers to form a more complex assembly. The robotic arm assembly3004may be utilized to move materials from one sterilized chamber to another through the connected transfer hatches3040.

FIG. 23shows an embodiment of an automatically controlled robotic arm within a sterilized three-dimensional printing chamber. The three-dimensional printing chamber and robotic arm control assembly3050comprises a sterilized chamber3052that contains at least one robotic arm assembly3060, a plurality of tubing that connects an automated control assembly3072with the robotic arm assembly3060, and a transfer hatch3082for the chamber. In this embodiment the sterilized chamber3052contains a printing assembly with a three-axis positioning gantry assembly3054and a printing head3056for dispensing material to a printing tray, which in this embodiment contains a multi-well plate3058. The sterilized chamber3052additionally contains a plurality of movable shelves3064attached to the walls of the sterilized chamber3052with supports3062for stacking a plurality of materials and/or printing trays, which in this embodiment contain a plurality of multi-well plates3066. In this embodiment the robotic arm assembly3060is utilized to move the empty multi-well plates3058to the printer assembly for printing with structural and biologically-active materials and to move those plates to the movable shelves3064for storage, incubation, and/or analysis. The automated control assembly3072utilizes a plurality of hydraulic and/or pneumatic tubing3068to move pressurized fluid into the actuators located on the robotic arm assembly3060to control the movements. The tubing lines are charged with sterile hydraulic and/or pneumatic fluid pressure through the filling assembly3070. In this embodiment an automated control assembly3072is utilized to control the movements of the robotic arm assembly3060. Pluralities of hydraulic and/or pneumatic pistons are arranged in a piston assembly3074that comprises pistons filled with fluid and the movements of the piston heads (not shown) are automatically controlled with a processing device3076. A memory storage device3078may store programs local to the automated control assembly3072to control the movements of the robotic arm within a program. The processing device3076may also process sensor data and alter the movements of the robotic arm assembly3060based on the programs stored within the memory storage device3078. The automated control assembly3072may additionally contain a wireless communication device to import robotic arm assembly control protocols, wireless sensor data, and/or communicate with an external network and/or a mobile device. The transfer hatch3082may be utilized to remove objects aseptically from the sterilized chamber3052and/or to connect to additional sterilized chambers to form a more complex assembly. The robotic arm assembly3060may be utilized to move materials from one sterilized chamber to another through the connected transfer hatches3082.

FIG. 24shows an embodiment of an automatically controlled robotic arm on a movable track within a sterilized three-dimensional assembly chamber. The three-dimensional chamber and robotic arm control assembly3100comprises a sterilized chamber3102that contains at least one robotic arm assembly3104, a plurality of tubing3110that connects an automated control assembly3116with the robotic arm assembly3104, and a transfer hatch3124for the chamber. In this embodiment the sterilized chamber3102contains a plurality of movable shelves3120attached to the walls of the sterilized chamber3102with supports3118for stacking a plurality of materials and/or printing trays, which in this embodiment contain a plurality of multi-well plates3122. In this embodiment the robotic arm assembly3104is utilized to move the stack multi-well plates3122on the movable shelves3120for storage, incubation, and/or analysis. For additional range of movement the robotic arm assembly3104may move along a track3106utilizing a wheeled or track assembly3108. The wheeled or track assembly3108may be powered by a hydraulic motor, a pneumatic motor, an electric motor, and/or a magnetic motor. The automated control assembly3116utilizes a plurality of hydraulic and/or pneumatic tubing to move pressurized fluid into the actuators located on the robotic arm assembly3104to control the movements. The tubing lines are charged with sterile hydraulic and/or pneumatic fluid pressure through the filling assembly3114. The transfer hatch3124may be utilized to remove objects aseptically from the sterilized chamber3102and/or to connect to additional sterilized chambers to form a more complex assembly. The robotic arm assembly3104may be utilized to move materials from one sterilized chamber to another through the connected transfer hatches3124.

FIG. 25shows an embodiment of an automatically controlled soft robotic arm within a sterilized three-dimensional assembly chamber. The three-dimensional chamber and soft robotic arm control assembly3150comprises a sterilized chamber3152that contains at least one soft robotic arm assembly3154, a plurality of tubing3170that connects an automated control assembly3174with the soft robotic arm assembly3154, and a transfer hatch3178for the chamber. In this embodiment the soft robotic arm assembly3154contains a base plate3156and a plurality of plates3158containing at least two inflatable sections, such as bags, bladders, or expandable reservoirs positioned accordingly within the plate assembly. As the inflatable sections in the plurality of plates3158are unevenly inflated3160, the soft robotic arm assembly3154bends towards a specific direction. The more inflatable sections available within the plurality of plates3158, the greater the degree of control for positioning the soft robotic arm assembly3154. The soft robotic arm assembly3154may contain a collar3162that separates the positioning section of the soft robotic arm from the gripping section3166of the robotic arm. The geometry of the plurality of plates3164in the gripping section3166may have a hinge in the center, where the inflatable sections cause the gripping section3166to close around an object. The automated control assembly3174utilizes a plurality of hydraulic and/or pneumatic tubing to move pressurized fluid into the actuators located on the soft robotic arm assembly3154to control the movements. The tubing lines are charged with sterile hydraulic and/or pneumatic fluid pressure through the filling assembly3172. The transfer hatch3178may be utilized to remove objects aseptically from the sterilized chamber3152and/or to connect to additional sterilized chambers to form a more complex assembly. The robotic arm assembly3154may be utilized to move materials from one sterilized chamber to another through the connected transfer hatches3178.

FIG. 26shows an embodiment of an automatically-controlled inflatable robotic arm within a sterilized three-dimensional assembly chamber. The three-dimensional chamber and inflatable robotic arm control assembly3200comprises a sterilized chamber3202that contains at least one inflatable robotic arm assembly3204, a plurality of tubing3228that connects an automated control assembly3232with the inflatable robotic arm assembly3204, and a transfer hatch3236for the chamber. In this embodiment the inflatable robotic arm assembly3204contains a rotating base plate3206, a movable hinge joint3208that positions the inflatable robotic arm assembly3204, a plurality of inflatable sections, which may be bags, bladders, or expandable reservoirs, including a first inflatable section3210and a second inflatable section3214, with a movable hinge section in between the two inflatable sections3210and3214. Additionally and/or alternatively the inflatable assemblies may be articulated along a seam and/or seal between two or more inflatable sections. A plurality of actuators3216pull cables3218, which are connected to the inflatable sections, and position the inflatable robotic arm assembly3204. An inflatable gripping section3224is available at the end of the inflatable arm assembly and closes around an object when pulled on by cable3222, which is controlled by actuator3220. The automated control assembly3232utilizes a plurality of hydraulic and/or pneumatic tubing3228to move pressurized fluid into the actuators located on the soft robotic arm assembly3204to control the movements. The tubing lines are charged with sterile hydraulic and/or pneumatic fluid pressure through the filling assembly3230. Pluralities of hydraulic and/or pneumatic pistons are arranged in a piston assembly3074that comprises cylinders filled with fluid and the movements of the piston heads (not shown) are automatically controlled with a processing device. The transfer hatch3236may be utilized to remove objects aseptically from the sterilized chamber3202and/or to connect to additional sterilized chambers to form a more complex assembly. The robotic arm assembly3204may be utilized to move materials from one sterilized chamber to another through the connected transfer hatches3236.

FIGS. 27A-27Eshow multiple embodiments of a sampling system which may provide periodic and/or continuous sampling of the materials located in the printing tray.

FIG. 27Ais a side view of a single-use printer chamber4000with a single-use aseptic sampling connector4008. The single-use printer chamber4000contains a printing tray4002, which in this embodiment is a multi-well tray. Each of the multi-well printing tray4002wells contains an opening, a seal, septum, and/or a closing assembly (not shown) that can allow material, particularly fluid-based materials, into a sampling tubing assembly4004below the printing tray4002. The sample tubing assembly4004may be aseptically connected to at least one external sanitary connection assembly4006. The sample tubing assembly4004may be expertly arranged to provide at least one sampling port for each well of the multi-well plate printing tray4002, may provide multiple sampling ports for each well, and/or may provide sampling ports to at least one representative well. The external sanitary connection assembly4006aseptically connects to an aseptic sampling connector4008, which in this embodiment is a TakeONE® aseptic sampling assembly. The operator may manually depress at least one of the spring-loaded thumb press actuators4010to push a needle through a self-sealing platinum-cured silicone septa (not shown), which provides a channel for the fluid material to pass through the single-use aseptic sampling connector4008and into a sampling collection container4012, which may be a bag, bottle, centrifuge tube, or other sterile container/connector type. The operator aseptically removes the sampling collection container4012utilizing a Quickseal® cutting tool (not shown) on the Quickseal® collar4014. The operator may use the sample from the sampling collection container4012originating from the single-use printer chamber4000printing tray4002in at least one external measurement device (not shown), to make a determination about the state of the material inside of the printing tray4002. Measurement results from the at least one external measurement device (not shown) may be manually entered and/or automatically (via a wired and/or wireless connection) communicated to the single-use printer chamber4000controller (not shown).

FIG. 27Bis a side view of a single-use printer chamber4030with an external continuous sampling system4040. The single-use printer chamber4030contains a printing tray4032, which in this embodiment is a multi-well tray. Each of the multi-well printing tray4032wells contains an opening, a seal, septum, and/or a closing assembly (not shown) that can allow material, particularly fluid based materials, into a sampling tubing assembly4034below the printing tray4032. The sample tubing assembly4034may be aseptically connected to at least one external sanitary connection assembly4036. The sample tubing assembly4034may be expertly arranged to provide at least one sampling port for each well of the multi-well plate printing tray4002, may provide multiple sampling ports for each well, and/or may provide sampling ports to at least one representative well. The external sanitary connection assembly4036aseptically connects to a tubing assembly4038, which provides the fluid material to at least one external continuous sampling system4040, which in this embodiment is a BioPAT® Trace analysis system. The external continuous sampling system4040may be a manual system, where an operator determines when a sample should be manually taken, or may be an automated system, where samples are taken at predetermined time intervals, by performing continuous sampling, and or by performing sampling based on intervals determined by the measurement data results. The fluid material tested by the external continuous sampling system4040may be read by single-use sensors and the fluid material may be discarded after testing through tubing line4042, which may allow the fluid material to exit the system via a drain (not shown) or by filling a carboy or container (not shown) for discarding after the run has completed. Additionally and/or alternatively the fluid material may be aseptically returned to the printing tray4032after testing. Measurement results from the at least one external continuous sampling system4040may be manually entered and/or automatically (via a wired and/or wireless connection) communicated to the single-use printer chamber4030controller (not shown).

FIG. 27Cis a side view of a single-use printer chamber4050with an internal sampling system. The single-use printer chamber4050contains a printing tray4052, which in this embodiment is a multi-well tray. An internal sampling system4054to provide measurement and sensor data for each of the individual wells of a multi-well tray may be incorporated within the printing tray4052itself. The internal sampling system4054incorporated into the printing tray4052may collect samples, measurements, and/or test data from the material inside of the printing tray4052. The internal sampling system4054incorporated into the printing tray4052may be integrated into the printing tray4052and/or multi-well plate and may be discarded after use (single-use). Alternatively, the internal sampling system4054incorporated into the printing tray4052may be removable from the printing tray4052and/or multi-well plate and may be inserted into an alternate printing tray4052and/or multi-well plate and re-sterilized for re-use (multi-use). The multi-use internal sampling system4054incorporated into the printing tray4052may incorporate some single-use elements such as single-use sensors, which may be discarded prior to inserting it into a new printing tray4052and/or multi-well plate. The internal sampling system4054incorporated into the printing tray4052may communicate the measurement and/or sensor data via a wired and/or wireless connection to the single-use printer chamber4050controller (not shown) and/or external measurement devices (not shown). Additionally and/or alternatively the material from the printing tray4052and/or multi-well plate may be sampled utilizing an internal sampling system4058incorporated into the printer chamber4050. Each of the multi-well printing tray4052wells contains an opening, a seal, septum, and/or a closing assembly (not shown), which can allow material, particularly fluid based materials, into a sampling tubing assembly4056below the printing tray4052. The sample tubing assembly4056may connect to at least one internal sampling system4058incorporated into the printer chamber4050. The internal sampling system4058incorporated into the printer chamber4050may be a manual system, where an operator determines when a sample should be manually taken, or may be an automated system, where samples are taken at predetermined time intervals, by performing continuous sampling, and or by performing sampling based on intervals determined by the measurement data results. The fluid material tested by the internal sampling system4058incorporated into the printer chamber4050may be read by single-use sensors and the fluid material may be discarded after testing (not shown) or may be returned to the printing tray4052after testing. Measurement results from the at least one internal sampling system4054incorporated into the printing tray4052and/or at least one internal sampling system4058incorporated into the printer chamber4050may be manually entered and/or automatically (via a wired and/or wireless connection) communicated to the single-use printer chamber4050controller (not shown).

FIG. 27Dis a top view of a multi-well plate printing tray4082on a printing platform4080, where each of the wells contains a sampling assembly4084, which may be an opening, a seal, a septum, and/or a closing assembly that can allow material, particularly fluid-based materials, into a sampling tubing assembly (not shown) below. The sampling assembly4084may be expertly arranged to provide at least one sampling port for each well of the multi-well plate printing tray4082, may provide multiple sampling ports for each well, and/or may provide sampling ports to at least one representative well. A self-healing septum may be pierced by a plurality of needles stored within the printing platform4080or within the sampling tubing assembly (not shown) below to extract a metered sample from the well in the printing tray4082.

FIG. 27Eis a top view of a multi-well plate printing tray4092on a printing platform4090, where each of the wells contains a plurality of single-use sensors4094. The single-use sensors may be expertly arranged to provide at least one single-use sensor4094for each well of the multi-well plate printing tray4092, may provide multiple single-use sensors4094for each well, and/or may provide at least one single-use sensors4094to at least one representative well.

FIG. 28shows an embodiment of a monitoring assembly for optical examination of the material in the printing tray. The single-use printer chamber4070contains a printing tray4072, which may be a multi-well plate, after structural material and/or biological material has been dispensed into the printing tray4072from the plurality of printing heads4074. In addition or alternatively to a printing head4074placed in a static position through a port within the single-use printer chamber4070, an optical measurement device4076, such as a microscopic camera, may be placed and utilized to optically measure the contents of the printing tray4072. A positional gantry aided by a positioning system4078comprising an array of cameras may be utilized to position the printing tray4072into the correct position to examine a specific section or well from the printing tray4072, wherein the printing tray4072may be positioned to reach the correct focal distance for viewing the contents of the printing tray4072. The optical measurement device4076, such as a microscopic camera, may contain a zoom lens and/or autofocus to provide a clear image of the contents. Lighting may be provided below by LED lights in the printing platform shining through the printing tray4072, or provided by an external source on the side of or below the single-use printer chamber4070. Additionally and/or alternatively the array of cameras utilized for the positioning system4078may be utilized for recording the contents of the printing tray4072, and not only for the positioning of the gantry setup. The array of cameras utilized for the positioning system4078may be positioned externally to the single-use printer chamber4070and view the internal assembly through a plurality of transparent windows and/or material. The optical measurement device4076and/or the array of cameras utilized for the positioning system4078may optically examine, measure, record, and store the optical data from the printing tray4072over a time period to determine and measure changes within the printed products inside of the printing tray4072, such as in a time-lapse growth profile. The optical measurements may be utilized to determine if a structural support is of sufficient quality prior to the addition of biologically active products. The optical measurements may also be utilized to determine the profile, density, coverage, adherence, invasiveness, health, and viability of cell growth onto a structural support. The optical measurements may additionally examine other factors, like color change from a chromic die for measuring pH, temperature, or other factors. The optical measurements may additionally be able to examine the printing tray4072contents for potential contamination of bacteria, fungi, viruses, or other unwanted cell growth.

FIGS. 29A-29Fshow a vacuum-forming unit internal to a sterile chamber for the manufacturing of components for use with biologically active materials.

FIG. 29Ashows a side view of a sterile chamber assembly5000for the manufacturing of components for use with biologically active materials. A sterile chamber5002, which may be made from flexible and/or rigid materials, contains at least one internal vacuum-forming unit5004for the forming of reproducible shapes utilizing vacuum-former support structure5006and a plurality of plastic sheets5008that are individually heated to a forming temperature using a heating element5014. The forming is achieved by overlaying the plastic sheet over a mold5020and activating a vacuum pressure on a plate with a plurality of holes5022through an external source on the vacuum assembly5024, which shapes the heated plastic sheet5008over the mold5020until it cools and solidifies in place. In this embodiment the vacuum-former support structure5006holds a plurality of plastic sheets5008above a sliding heater element assembly5012. When the sliding heater element assembly5012is in the exterior position on a sliding platform5018, a single plastic sheet5010is released and falls into place using guides into the heating tray holder. The sliding heater element assembly5012contains the heating element5014and an insulating layer5016, which prevents the heat from the heating element5014to heat the plurality of plastic sheets5008above and make them reach forming temperature, or to protect the sterile chamber5002itself while heating the single plastic sheet5010below. The heating element5014may be an electrical heater, a single-use chemical reaction heating device, or may operate through the circulation of an externally-heated fluid source such as heated water, glycol, and/or steam. The plurality of plastic sheets5008may have a low melting-point profile so as not to require high heating temperatures for forming structures. The vacuum assembly5024may connect to an external vacuum source through the sterile chamber5002. The vacuum assembly5024may utilize an aseptic connection and/or a sized air filter (not shown), such as a sterilizing grade air filter (not shown), which would allow sufficient airflow to result in the vacuum pressure required to shape the heated plastic sheet5010around the mold5020. The sterile chamber5002may additionally contain a robotic arm5026for the placement of the mold5020, for the cutting and/or die-cut punching of the formed product, for the removing of the formed product from the vacuum-forming unit5004, for the transfer of the formed product to a storage assembly (not shown) and/or to a connected sterile chamber (not shown) through a transfer hatch5034, and for the removal of waste plastic material from the vacuum-forming unit5004into a waste container (not shown). The robotic arm may contain a cutting assembly such as a cutting blade (not shown) or a die-cut assembly (not shown) within a robot-arm transfer head5028for cutting out the formed product after the vacuum forming has been completed. The robot-arm transfer head5028may additionally contain a vacuum assembly for creating a seal and holding onto the formed product during the transfer process. The mold5020may be a 3D printed product from another part of a sterile multi-chamber assembly (not shown). The sterile chamber5002may additionally contain a vent filter5030, such as a sterilizing-grade vent filter, of sufficient size to maintain the integrity of the rigid and/or flexible sterile chamber5002. For example the vent filter5030may have a large size if a flexible chamber is utilized, in order to prevent the chamber from collapsing in on itself during the maximum vacuum force from the vacuum assembly5024. An additional support skeleton may be utilized to strengthen a flexible sterile chamber5002. An additional transfer hatch support structure5032may be utilized to strengthen the transfer hatch5034during the maximum vacuum force from the vacuum assembly5024. The transfer hatch support structure5032may be in a closed state during vacuum pressure inside the sterile chamber5002to prevent leakage of the vacuum force to an externally connected chamber (not shown) connected to the transfer hatch5034.

FIG. 29Bshows a side view of a sterile chamber5002containing at least one internal vacuum-forming unit5004. In this embodiment the sliding heater element assembly5012′ is in the internal position on the sliding platform5018. The heating element5014′ is engaged in covering and heating the single plastic sheet5010(not shown), while the insulating layer5016is preventing the heat from the heating element5014′ from heating the plurality of plastic sheets5008above the assembly. In preparation for the vacuum force, the transfer hatch support structure5032is engaged into the closed state to strengthen the transfer hatch5034and prevent leakage of the vacuum force to an externally connected chamber (not shown).

FIG. 29Cshows a side view of a sterile chamber5002containing at least one internal vacuum-forming unit5004. In this embodiment the sliding heater element5014has heated the single plastic sheet5010′ to the proper forming temperature. The vacuum assembly5024′ is activated using an external vacuum source on the vacuum former, providing vacuum pressure on a plate with a plurality of holes5022. The heated single plastic sheet5010′ is released and follows the guides to drop directly over the mold5020′. As it covers the mold5020′, the vacuum pressure from below stretches the single plastic sheet5010′ to take the shape of the mold and adhere to the plate with a plurality of holes5022. A frame (not shown) may or may not be used to hold the edges of the plastic sheet5010′ in place during the vacuum-forming process. The vent filter5030′ is sized to provide sufficient airflow into the sterile chamber5002during the maximum vacuum force from the vacuum assembly5024′ when it is engaged.

FIG. 29Dshows a side view of a sterile chamber5002containing at least one internal vacuum-forming unit5004. The vacuum pressure from the vacuum assembly5024is stopped and the formed single plastic sheet5010′ around the mold5020is allowed sufficient time to cool and solidify. In this embodiment the robotic arm5026′ moves to cover the formed product (not shown) from the single plastic sheet5010′ over the mold5020. The robot-arm transfer head5028contains a die-cut assembly (not shown) for cutting out the formed product after the vacuum forming has been completed. The robot-arm transfer head5028additionally contains a vacuum assembly for creating a seal and holding onto the formed product during the transfer process to a storage area. In this embodiment the formed product is a multi-well plate for use with a printing tray and printing of structural materials and biologically-active materials within a three-dimensional printing chamber that is connected directly or indirectly to the sterile chamber5002through the transfer hatch5034.

FIG. 29Eshows a side view of a sterile chamber5002containing at least one internal vacuum-forming unit5004. The sliding heater element assembly5012slides into the external position on the sliding platform5018and an additional single plastic sheet5036is released and falls into place using guides into the heating tray holder. The robotic arm5026moves into position to remove the remaining plastic material5010′ on the vacuum former plate with a plurality of holes5022. The robot-arm transfer head5028utilizes an internal vacuum and/or plurality of suction cups to attach to the remaining plastic material5010′, lifts it over the mold5020, and places it into a waste container (not shown) inside of the sterile chamber5002for discarding. The transfer hatch support structure5032is opened to allow for the formed product to be moved through the transfer hatch5034into another chamber, where it may be utilized or undergo further processing.

FIG. 29Fshows a side view of a sterile chamber5002containing at least one internal vacuum-forming unit5004. The vacuum-forming unit5004is returned to the original state for forming an additional product from a new single plastic sheet5036. The use of a vacuum-forming unit internal to a sterile assembly allows to form a product of a particular design by re-using a mold so that a plurality of products formed in such manner, such as a multi-well assembly for use as a printing tray, may be utilized over and over again within a process inside a network of sterile chamber5002assemblies. A vacuum-forming process is faster, easier and has a more consistent manufacturing than current three-dimensional printing methods.

FIGS. 30A-30Eshow an injection-molding unit internal to a sterile chamber for the manufacturing of components for use with biologically active materials.

FIG. 30Ashows a side view of a sterile chamber assembly5050for the manufacturing of components for use with biologically active materials. An internal sterile chamber5054, which may be made from flexible and/or rigid materials, contains at least one internal injection-molding unit5062for the forming of reproducible shapes utilizing a melted material injected into a solid mold5078. In this embodiment the sterile chamber assembly5050contains an outer jacketed container5052where a cooling fluid, such as cold sterile filtered water, is recirculated5056to maintain a stable internal chamber temperature with the heating element5070from the internal injection-molding unit5062. A plurality of plastic supports (not shown), with openings to promote fluid circulation, may be in place between the outer jacketed container5052and the internal sterile chamber5054to hold the internal sterile chamber5054in place. To connect the internal sterile chamber5054with the outer jacketed container5052at least one vent filter assembly5060utilizes at least one connection tube5058. To connect the internal sterile chamber5054with the outer jacketed container5052a transfer hatch assembly5088utilizes a connection tube to an internal transfer hatch5086. The at least one internal injection-molding unit5062inside of the internal sterile chamber5054contains a stand5064, a hopper5066filled with a low-melting-point moldable material, such as plastic pellets, and a feed tube5068to feed the moldable material from the hopper5066into the insulated heating element5070. The insulated heating element5070heats the moldable material, such as the plastic pellets, into a melted material which is extruded under compression force utilizing a compression motor5074. The melted material is extruded utilizing a metered dispenser5076into a solid mold5078. As heat rises from the insulated heating element5070inside of the internal sterile chamber5054, an insulated thermal barrier5072may be used to prevent deformation and/or damage of the internal sterile chamber5054walls from the excess heat. In addition to fluid circulation in the jacketed container5052, to maintain a stable temperature the vent filter assembly5060and/or a plurality of vent filters may be utilized to perform a fluid air exchange to remove excess heat from the internal sterile chamber5054. Both the jacketed fluid and/or the air exchange may utilize an external heat exchanger device (not shown) to maintain and control the internal temperature of the internal sterile chamber5054. A plurality of temperature sensors (not shown) may be located within the internal sterile chamber5054and/or the outer jacketed container5052to provide temperature measurements for the proper regulation of temperature by external devices (not shown). The solid mold5078may comprise at least two parts that are closed and sealed with a motor5080. The injection molded material is allowed to cool and solidify within the solid mold5078and is removed utilizing a removal tool5082which places the final injection molded product into a holding container5084.

FIG. 30Bshows a side view of an internal sterile chamber5054containing at least one internal injection-molding unit5062′. In this embodiment the insulated heating element5070′ is actively heating the moldable material from the hopper5066, such as the plastic pellets, into a melted material. The insulated heating element5070′ may be an electrical heater, a single-use chemical reaction heating device, or may operate through the circulation of an externally-heated fluid source such as heated water, glycol, and/or steam. The compression motor5074′ exerts a force on the melted material and pushes it through the metered dispenser5076into the solid mold5078′. The compression motor5074′ may be driven by hydraulic, pneumatic, electric, or magnetically controlled methods (not shown). The injection-molded material is allowed to cool and solidify within the solid mold5078′.

FIG. 30Cshows a side view of an internal sterile chamber5054containing at least one internal injection-molding unit5062″. In this embodiment at least one part of the solid mold5078″ is separated by retracting the first part using the motor5080′, thereby exposing the final injection-molded product5090.

FIG. 30Dshows a side view of an internal sterile chamber5054containing at least one internal injection-molding unit5062′″. In this embodiment the removal tool5082′ is activated and pushes, pulls, and/or scrapes the final injection-molded product5090′ out of the second part of the solid mold5078and into a holding container5084.

FIG. 30Eshows a side view of an internal sterile chamber5054containing at least one internal injection-molding unit5062. In this embodiment the internal injection-molding unit5062has returned to the original state in that the compression motor5074has returned to the original position. As the internal barrel (not shown) of the compression motor5074moves up the plastic pellets of the moldable material from the hopper5066′, it fills the empty chamber (not shown) causing a reduction in height of the moldable material from the hopper5066′ reservoir. The motor5080can return the first part of the solid mold5078to the second part forming a sealed container for the next injection mold. The final injection-molded product5090′ may be individually removed from the holding container5084utilizing a robotic arm (not shown) or the entire holding container5084may be removed after it has been filled with a plurality of final injection-molded products5090′. The final injection molded product5090′ may be moved through the internal transfer hatch5086and the outer transfer hatch5088into another chamber where it may be utilized or undergo further processing.

FIGS. 31Aand B show a laser-cutting device internal to a sterile chamber for the manufacturing of components for use with biologically active materials.

FIG. 31Ashows a side view of a sterile chamber assembly5100for the manufacturing of components for use with biologically active materials. A sterile chamber5102, which may be made from flexible and/or rigid materials, contains at least one laser-cutting device5104for the precision-cutting of shapes utilizing a laser5114to cut material from a substrate5118. In this embodiment the at least one laser-cutting device5104is supported by a solid base5106and a two-axis controller positioning system. In this embodiment the laser-cutting device5104contains a platform5108that moves along the horizontal x-axis along a track5126. The laser assembly5114is held by a support structure5110, which contains a track5112for the z-axis movements of the laser assembly5114. The laser beam5116comes from the assembly and is utilized to cut through a substrate5118located on the platform5108with precision. The laser cut products5120from the laser assembly may be utilized for manufacturing components for use with biologically active materials. Additionally and/or alternatively the laser may be utilized to laser-label some materials for information and/or coding. A plurality of vent filters5122may be utilized to regulate the pressure of the sterile chamber5102and may additionally be utilized to remove smoke or aerosolized particulates from the laser5116burning through the substrate5118. The exterior wall of the sterile chamber5102may be coated with an absorbing material to block and reduce the intensity of the laser5116at the specific wavelengths that the laser assembly5114operates at. This will reduce the risk of exposure to the operator even if they are not wearing laser safety eyewear for protection. The laser-cut products5120from the substrate5118may be moved out of the sterile chamber5102through a transfer hatch5124. The internal laser assembly5114may be powered by an external electrical connection, an internal battery storage mechanism, an inductively-charged electrical connection, a chemical reaction, a microwave or visual line-of-sight power source, or other wireless power source. The two-axis motor control mechanism may be driven by hydraulic, pneumatic, electric, or magnetically controlled methods (not shown).

FIG. 31Bshows a side view of a sterile chamber5102that contains at least one laser-cutting device5104for the cutting of shapes utilizing a laser5114to cut material from a substrate5118. In this embodiment the at least one laser-cutting device5104′ contains a platform5108′ which has moved along the horizontal x-axis via a track5126. The laser assembly5114has moved along the z-axis via a track5112. The combined movements of the laser assembly5114and of the platform5108′ were able to control the laser beam5116to precisely cut the laser-cut products5120from the substrate5118located on the platform5108′. Alternatively the laser assembly5114may be in a fixed position inserted through a port (not shown) in the sterile chamber5102and the platform5108may move along two-axis for positioning the substrate5118. Additionally the laser assembly5114may be aseptically connected to the sterile chamber5102or sterilized in place during connection with the sterile chamber5102as previously described. The final laser-cut products5120may be individually removed from the platform5108′ and placed into a holding container (not shown) utilizing a robotic arm (not shown). Additionally or alternatively a robotic arm (not shown) may be utilized to assemble the final laser-cut products5120into an assembly (not shown). The robotic arm (not shown) may be utilized to move the individual laser-cut products5120or the assembled product (not shown) into another chamber where it may be utilized or undergo further processing.

FIG. 32shows an ultrasonic-welding device internal to a sterile chamber for the joining of manufactured components for use with biologically active materials. A sterile chamber5152, which may be made from flexible and/or rigid materials, contains at least one ultrasonic-welding device5154which welds at least two separate materials, such as two plastic materials, together to form a single component. In this embodiment the at least one ultrasonic-welding device5154may contain a platform base5156, which may contain a vibrational damper (not shown) or be made from vibrational-damping materials to prevent the vibrational energy from the ultrasonic welder from being transferred to the sterile chamber5152. The ultrasonic-welding device5154may contain a movable platform5158on the platform base5156. The movable platform5158may contain a support structure5160to hold an ultrasonic-welding assembly5162in place above an anvil5182. The ultrasonic-welding assembly5162may contain a piston5164, which may be driven by hydraulic, pneumatic, electric, or magnetically controlled methods (not shown). The ultrasonic-welding assembly5162may contain a transducer5166, a converter5168, a booster5170, a collar5172, a sonotrode5174, and a horn5176. The sonotrode5174and horn5176assembly comes in contact with a first weldable component5178that is in physical contact with a second weldable component5180. The first weldable component5178and the second weldable component5180are lined up for ultrasonic welding on the anvil5182. Electrical energy is transformed into vibrational energy in the ultrasonic-welding assembly5162and makes the interface between the weldable components5178and5180vibrate at thousands of times per second. The vibrations create frictional heat that causes the materials from the weldable components to melt together forming a single welded component. The standard frequencies utilized with ultrasonic sonotrodes range from about 20 kHz to about 70 kHz and the amplitude of the vibrations is approximately 13 to 130 micrometers. A robotic arm (not shown) within the assembly may precisely position the first weldable component5178and the second weldable component5180. The ultrasonic-welding device5154may be powered by an external electrical connection, an internal battery storage mechanism, an inductively-charged electrical connection, a chemical reaction, a microwave or visual line-of-sight power source, or other wireless power source. The robotic arm (not shown) may additionally be utilized for removing the welded assembly from the anvil5182and placing it into a holding container (not shown) or moving the welded assembly into another chamber through a transfer hatch5186where it may be utilized or undergo further processing.

FIG. 33shows a three-dimensional printing assembly inside of a sterile single-use chamber6000, which may be made from flexible and/or rigid materials and contains a plurality of printing heads6002, which are used to dispense materials into a printing tray. In this embodiment the printing platform is a multi-segmented tile printing platform that contains a plurality of printing trays6004on a plurality of multi-segmented tiles, where at least one of the tiles is missing to allow for each of the individual tiles to move to all possible positions on the printing platform. The multi-segmented printing platform may be driven by hydraulic, pneumatic, electric, or magnetically-controlled methods (not shown) or as previously described. In this embodiment the multi-segmented printing platform is missing at least two tiles6006and6008to properly position the central tile in both the x-axis and z-axis during the printing process with a plurality of fixed printing heads6002. In this embodiment the plurality of printing trays6004comprises multi-well plates, wherein the plurality of printing heads6002may dispense a metered amount of liquid media, three-dimensionally-print structural material, and/or three-dimensionally-print cells onto the structural material, and dispense a metered amount of at least one drug product for the in-vitro testing of the cell products inside of the multi-well plates. The applications could be preclinical efficacy and toxicology testing of drug products on the cells or other bioactive materials. A plurality of cell types may be utilized, for example in a method for screening a patient's own cells grown up in a bioreactor and determine the optimal drug treatment regimen based on the efficacy and toxicology measured within the plurality of printing trays6004. The sterile single-use chamber6000may be regulated to a temperature for incubating the cells within the plurality of printing trays6004. A transfer hatch6010may be utilized for the aseptic transfer of the materials inside of the sterile single-use chamber6000or for the transfer of materials to a network of chambers connected by the transfer hatch6010.

FIG. 34shows a three-dimensional printing assembly inside of a sterile single-use chamber6020, which may be made from flexible and/or rigid materials and contains a plurality of printing heads6026that are used to dispense materials into a printing tray6030. In this embodiment the printing tray6030moves along a printing platform (not shown) along a three-axis gantry setup6028during the printing process with a plurality of fixed printing heads6026. In this embodiment the printing tray6026is a multi-well plate, where the plurality of printing heads6026may dispense a metered amount of liquid media, three-dimensionally-print structural material, and/or three-dimensionally-print cells onto the structural material, and dispense a metered amount of at least one drug product for the in-vitro testing of the cell products inside of the multi-well plates. In this embodiment a plurality of robotic arms6032and6038may be utilized to place an empty printing tray6036onto the printing platform (not shown) for printing with the plurality of printing heads6026. The robotic arms6032and6038may additionally be utilized to move a printing tray6040, which has completed printing, to a storage location. The storage location may contain a plurality of moving racks for stacking the printing trays within the sterile single-use chamber6020to maximize the use of the space as well as to increase the throughput of printing trays6040capable of being printed, measured, studied, and/or screened within the chamber6020. In this embodiment the plurality of robotic arms6032and6038move along a plurality of tracks6034to increase the range of motion for the robotic arms6032and6038and increase the area available for stacking and/or storage of printing trays6036and6040. The sterile single-use chamber6020may be regulated to a temperature for incubating the cells and/or bioactive products within the plurality of printing trays6004. In this embodiment the sterile single-use chamber6020is a jacketed chamber that contains an outer jacketed chamber6022and uses the circulation6024of a fluid, such as sterile filtered water, for regulating and/or maintaining the proper temperature for the chamber6020. A transfer hatch (not shown) may be utilized for the aseptic transfer of the materials inside of the sterile single-use chamber6020or for the transfer of materials to a network of chambers connected by the transfer hatch (not shown).

FIG. 35shows a top view of a network of single-use chambers, each with their own functionalities, which forms a single-use manufacturing assembly operating within a sterile environment. The network of single-use chambers6050provides a method to connect individual single-use chambers within a certain arrangement to perform a manufacturing, testing, screening, and/or measurement function. In this embodiment the central single-use three-dimensional printing chamber6052is connected to a plurality of other chambers each with their own functionalities. In this embodiment the process starts in the single-use vacuum-forming chamber6058, which utilizes a vacuum-forming unit6062to form a heated plastic piece over a mold by exploiting a strong vacuum using the external vacuum port6064. In this embodiment the vacuum-formed product is a multi-well plate6060. A robotic arm6066grabs the formed multi-well plate6060and places it onto a conveyor belt6068that moves the multi-well plate6060through a transfer hatch6072, when the vacuum is not running and a hatch support structure6070is in the opened state, and into a single-use multi-well-plate coating chamber6074. The conveyor belt may be made from sterilizable materials and may be driven by hydraulic, pneumatic, electric, or magnetically-controlled methods (not shown) or as previously described. Alternatively or additionally the multi-well plates or other material inputs may be pre-sterilized in an alternate sterile chamber (not shown) and connected to the network of single-use chambers6050. Alternatively the multi-well plates or other material inputs may be aseptically connected via a transfer hatch (not shown). In another embodiment the multi-well plates or other inputs may utilize a sterilization chamber (not shown) where the non-sterile materials are inserted and the container is sealed, the chamber is sterilized utilizing a validated method and load configuration such as by chemical sterilization by vaporized hydrogen peroxide (VHP) and the sterilization chamber is either connected to the network of single-use chambers6050by an aseptic connector (not shown) or was already connected and the barrier (not shown) between the connected chambers is aseptically removed. An additional conveyor belt6076moves the multi-well plate6080below a fixed printing head6078that spray-coats the multi-well plate6080in preparation for the printing process. The spray coating may prepare the multi-well plate6080for the adhesion of structural materials and/or cells, or may be utilized to keep the cells suspended and not adhering to the side walls. Additionally and/or alternatively the spray coating may add growth factors that support the growth of certain cell types within the multi-well plate6080. The spray coating from the printing head6078may be non-specific and coat the entire plate or may be specific and coat only specific regions of the plate or with a specific pattern or design. The completed multi-well plate6054after spray coating moves along a conveyor6082through a transfer hatch6084and into the central single-use three-dimensional printing chamber6052. The multi-well plate6054enters a printer three-axis gantry setup (not shown) and undergoes three-dimensional printing with a plurality of printing heads6052. In this embodiment the plurality of printing heads6052may dispense a metered amount of liquid media, three-dimensionally-print structural material, and/or three-dimensionally-print cells onto the structural material, and dispense a metered amount of at least one drug product for the in-vitro testing of the cell products inside of the multi-well plates. The printed multi-well plate6054moves along a conveyor belt6056through a transfer hatch into a single-use incubation and storage chamber6100. The printed multi-well plate6054moves onto a conveyor belt6106, where the robotic arm6110moving along a track grabs it and places it in an open storage space inside the stackable incubation chamber. In this embodiment the single-use incubation and storage chamber6100is a jacketed chamber that contains an outer jacketed chamber and uses the circulation6102of a fluid, such as sterile filtered water, for regulating and/or maintaining the proper temperature within the chamber. The multi-well plates may be tracked by an individualized labeled barcode (not shown). After a period of time for incubation, the multi-well plates6108and6112inside of the single-use incubation and storage chamber6100are sent for testing and examination. The robotic arm grabs the incubated multi-well plate, such as6108, and transfers it to the conveyor belt6106, which moves it to the central single-use three-dimensional printing chamber6052and down a conveyor belt6118through a transfer hatch into the single-use optical examination chamber6114. The conveyor belt6118loads the incubated multi-well plate into the three-axis gantry setup6122where it is positioned at the proper focal length from the optical measurement device6120, which is at a fixed position within the chamber. In this embodiment the optical measurement device6120is a microscopic imager, where the incubated multi-well plate is illuminated by LED lights (not shown) and the microscopic images are captured, stored on an external device, and compared to previous images. After the incubated multi-well plate has completed optical measurement in the single-use optical examination chamber6114, it is loaded back onto the conveyor belt6118through the central single-use three-dimensional printing chamber6052, through a transfer hatch6088, onto the conveyor belt6090, and into the single-use sampling chamber6086. The single-use sampling chamber6086contains a platform6092that utilizes a previously described sampling mechanism (not shown) to sample the material from the incubated multi-well plate. The sampled material moves through a sampling tubing assembly6094and to a single-use sampling assembly6096, which in this embodiment is a TakeONE® single-use sampling assembly. The material sampled from the incubated multi-well plate enters into a sterile container6098, which is aseptically removed from the single-use sampling assembly6096and tested on an external measurement device (not shown). After sampling has been completed, the incubated multi-well plate may be returned to the single-use incubation and storage chamber6100for further incubation, sent to the single-use three-dimensional printing chamber6052for further printing and/or the addition of media or a metered dose of at least one drug product, sent to the single-use optical examination chamber6114for additional optical examination, sent to a separate chamber (not shown) for non-incubated storage, or removed from the chamber and discarded (not shown). Alternatively all of the multi-well plates may be discarded when the batch testing has been completed and the plurality of single-use chambers6052,6058,6074,6086,6100, and6114are disconnected, sterilized for disposal, and discarded.

The following items may be useful for understanding the invention:

Item 1. A three dimensional printing device comprising:a sterilizable printer assembly includingat least one printing head,a printing platform, anda driving mechanism adapted to achieve a relative displacement between the at least one printing head and the printing platform along two or three degrees of freedom;a printer housing enclosing the printer assembly in a sterile manner,at least one aseptic connector fluidly connected to a corresponding one of the at least one printing head.

Item 2. The three dimensional printer of item 1, wherein a pre-sterilized printer head may be aseptically connected to the sterile three dimensional printing chamber.

Item 3. The three dimensional printer of item 1, wherein a non-sterilized printer head may be inserted into a printer head insertion assembly within the sterile three dimensional printing chamber and sterilized utilizing a chemical sterilization method.

Item 4. The printer head of item 1, wherein the printer head is utilized to dispense a metered volume of at least one drug product. The at least one drug product may be fluidly connected to the printer head via an aseptic connection.

Item 5. A printing system comprising:

a three dimensional printing device having:a sterilizable printer assembly includinga tethered multiaxis printing assembly includingat least one printing head, with a tubing line to a port in the chamber for transferring fluid and/or material inputs to be dispensed by the at least one printing head.an aseptic connector to connect an external fluid and/or material connection to the port in the sterile chamber.a plurality of soft robotics actuators for positioning the tethered multiaxis printing assembly into position over a fixed printing platform.an externally driven hydraulic and/or pneumatic drive mechanism to control the movements of the soft robotic actuators.a central body to connect the at least one printing head to the plurality of soft robotics actuators and the inputs and fluid connections for dispensing and positioning the assembly for printing on the printing platform.

Item 6. A printing system comprising:

a three dimensional printing device having:a sterilizable printer assembly includingan untethered multiaxis printing assembly includingat least one printing head, with a plurality of fluid reservoirs.a plurality of robotic appendages for positioning the untethered multiaxis printing assembly into position over a fixed printing platform.an internally powered drive mechanism to control the movements of the robotic appendages.a central body to connect the at least one printing head, the plurality of fluid reservoirs, and the plurality of robotic appendages for dispensing and positioning the assembly for printing on the printing platform.

Item 7. A robotic arm assembly comprising:a sterilizable chamber includinga robotic arm assembly made from sterilizable components internal to the sterilizable chamber includingat least one actuatorat least one articulating supportat least one gripping assemblyat least one control mechanismat least one positional systemand at least one driving mechanism adapted to achieve a relative displacement within the sterilizable chamber.

Item 8. The robotic arm assembly of item 7, wherein the at least one driving mechanism is a hydraulic and/or pneumatic fluid pressure source. The hydraulic and/or pneumatic fluid pressure source may originate from an external control assembly where the fluid lines are aseptically connected to the robotic arm assembly within the sterilized chamber.

Item 9. The robotic arm assembly of item 7, wherein the at least one control mechanism is a manual control mechanism where an operator may manually manipulate the controls of the robotic arm assembly internal to the sterile chamber.

Item 10. The robotic arm assembly of item 7, wherein the at least one control mechanism is an automated control mechanism which is controlled by a computer setup and/or an electronic device.

Item 11. The robotic arm assembly of item 7, wherein the at least one positional system is a plurality of cameras located on the sterile chamber and/or a plurality of cameras located on the robotic arm assembly.

Item 12. The robotic arm assembly of item 7, wherein the robotic arm internal to the sterile chamber is utilized for moving and stacking a plurality of components, materials, and or printer trays.

Item 13. The robotic arm assembly of item 7, wherein the robotic arm internal to the sterile chamber is utilized for assembling at least two components into an assembly.

Item 14. The robotic arm assembly of item 7, wherein the robotic arm internal to the sterile chamber is utilized for moving a plurality of components from one sterile chamber to another sterile chamber aseptically connected through the transfer hatches.

Item 15. The robotic arm assembly of item 7, wherein the robotic arm internal to the sterile chamber is a soft robotics assembly comprised of soft flexible materials and a plurality of inflatable sections.

Item 16. The robotic arm assembly of item 7, wherein the robotic arm internal to the sterile chamber is an inflatable robotics assembly comprised of a plurality of inflatable sections for the robotic arm supporting structures.

Item 17. A vacuum forming assembly comprising:a sterilizable chamber includinga vacuum forming assembly made from sterilizable components internal to the sterilizable chamber includingat least one rigid support structureat least one vacuum platform with a plurality of holesat least one vacuum pressure sourceat least one heating elementat least one dispensing mechanism for dispensing moldable plastic sheetsat least one mold,and at least one vent filter assembly.

Item 18. An injection molding assembly comprising:a sterilizable chamber includingan injection molding assembly made from sterilizable components internal to the sterilizable chamber includingat least one rigid support structureat least one reservoir with moldable materialat least one heating elementat least one compression sourceat least one dispensing unitat least one solid mold comprising of at least two partsand at least one vent filter assembly.

Item 19. A laser cutting assembly comprising:a sterilizable chamber includinga laser cutting assembly made from sterilizable components internal to the sterilizable chamber includingat least one rigid support structureat least one 2 axis positioning assemblyat least one laser assemblyat least one power sourceat least one substrateand at least one vent filter assembly.

Item 20. An ultrasonic welding assembly comprising:a sterilizable chamber includingan ultrasonic welding assembly made from sterilizable components internal to the sterilizable chamber includingat least one rigid support structureat least one ultrasonic welder comprising of a pistona transducera convertera boostera sonotrodea hornan anvilat least two materials two weld together.and at least one vent filter assembly.

Item 21. A single-use sampling assembly comprising:a sterilizable chamber includinga sampling assembly made from sterilizable components internal and external to the sterilizable chamber includingat least one fluid connection to the container to be sampledat least one single-use aseptic connection to the sterile chamberat least one sampling method for drawing fluid material from the desired chamber within the container to be sampledat least one collection container for holding the sampled fluid from the desired chamber within the container to be sampledat least one aseptic disconnection method for removal of the container containing the sample fluid.

Item 22. A continuous sampling assembly comprising:a sterilizable chamber includinga sampling assembly made from sterilizable components external to the sterilizable chamber includingat least one fluid connection to the container to be sampledat least one aseptic connection to the sterile chamberat least one sampling method for drawing fluid material from the desired chamberat least one external measurement deviceat least one pump and/or drive mechanism to draw the fluid to the at least one external measurement device.

Item 23. An internal sampling assembly comprising:a sterilizable chamber includinga sampling assembly made from sterilizable components internal to the sterilizable chamber includingat least one fluid connection to the container to be sampleda sampling method for drawing fluid material from the desired chamberan internal measurement device.

Item 24. An optical measurement assembly comprising:a sterilizable chamber includingan optical measurement assembly made from sterilizable components internal and/or external to the sterilizable chamber includingat least one optical measurement device, preferably a microscopic camera, containing a plurality of lenses and an autofocus.at least one LED light source.and at least one memory storage device.

Item 25. The optical measurement assembly of item 24, wherein the optical measurement assembly is comprised of a plurality of cameras positioned externally to the sterile chamber and views the internal assembly through a plurality of transparent windows and/or transparent material.

Item 26. A plurality of printer trays comprising multi-well plates comprising a plurality of wells.

Item 27. The plurality of printer trays of item 26, wherein the multi-well plates may be manufactured inside of the sterile chamber utilizing the at least one three dimensional printer, the at least one vacuum forming unit, the at least one injection molding unit, the at least one laser cutting unit, and/or the at least one ultrasonic welding unit.

Item 28. The plurality of printer trays of item 26, wherein the printer trays may be incubated under temperature regulated conditions for the growth of cells in nutrient rich media within the trays or within the scaffolding of the manufactured products.

Item 29. At least two sterile chambers may be aseptically connected together via the transfer hatch connections to form a network of sterile chambers where each sterile chamber performs a specific task.

Item 30. The network of sterile chambers of item 29, wherein the network of sterile chambers includes the manufacturing of a plurality of multi-well plates and spray coating with bioactive materials for growing cells in a nutrient rich media under incubated conditions, the use of cells grown in multi-well plates for the screening of three dimensional printed cell products, in vitro efficacy and toxicity studies of the effects of metered volumes of at least one drug products on three dimensional printed incubated cells within the multi-well plates.

Item 31. The network of sterile chambers of item 29, wherein the network of sterile chambers includes the manufacturing of a plurality of multi-well plates and spray coating with bioactive materials for growing cells in a nutrient rich media under incubated conditions, the use of cells grown in multi-well plates for the screening of three dimensional printed cell products, in vitro efficacy and toxicity studies of the effects of metered volumes of at least one drug products on three dimensional printed incubated cells within the multi-well plates.

Item 32. The network of sterile chambers of item 29, wherein the network of sterile chambers includes the manufacture of biologically active cell products, the printing of scaffolding materials for coating with bioactive products, the manufacture of complex multi-material assemblies for coating with bioactive products, the manufacture of complex multi-material medical devices with bioactive coatings, the printing of cells and cell products onto complex multi-material assemblies.

Item 33. The network of sterile chambers of item 29, wherein the network of sterile chambers includes the printing of cell products, cellular structures, scaffoldings, organs, and organ simulants grown from an individual patient's cells in a single-use bioreactor, processed, dispensed, incubated, and grown in a single-use sterile environment. The biologically grown product may be aseptically removed from the assembly and implanted into a patient in appropriate health care facility.

Item 34. The network of sterile chambers of item 29, wherein the network of sterile chambers includes the manufacture of biologically active products on biosensors inside a network of sterile chambers includes the printing of biologically active products onto electronic devices and/or substrates to add to electronic devices for the detection of an analyte. The analyte may be detected through the combination of a biological component with a physicochemical detector.

Item 35. The network of sterile chambers of item 29, wherein the network of sterile chambers includes the manufacture of biologically active products on diagnostic membranes inside a network of sterile chambers includes the printing of biologically active products onto a diagnostic membrane. The diagnostic membrane with the biologically active product may be assembled with a protective covering and/or a device delivery tool for the diagnostic testing, reading of results, and test analysis.

Item 36. The network of sterile chambers of item 29, wherein the network of sterile chambers includes the manufacture of custom filters by printing biologically active products onto filter membranes which are assembled into a completed filtration unit. Bioactive materials such as antibodies may be utilized with a custom filter membrane for the capture and removal or for the capture and elution of a specific material from a filtered fluid.