Instrumentality for sequestering liquids based on density:method and apparatus

An apparatus and method for collecting whole blood and then separating it into components for subsequent use or storage. A self-contained bag set is used to collect the sample, which may then be placed into a device adapted to fit into a centrifuge for separation of components. Each component is then sequentially extracted according to density, with a sensor present in the device to control the operation of valves directing the collection of each component. The sensor may be reading one or more of the following characteristics: infrared, optics, density, weight, radioactive, fluorescence, color, magnetism, ultrasonic, capacitance wherein the characteristic is inherent in the blood and blood component or is an additive. Each component may then be separated into its own storage container. The preferred sensors include optics and weight. Besides blood density separation, the device may contain a solution including cells, proteins, subcellular particles or viruses which may be mixed with affinity media or antibodies prior to separation.

CROSS-REFERENCE TO RELATED APPLICATION

This application is a national stage application based on PCT application no. PCT/US2005/029288 filed on. Aug. 16, 2005, which claimed priority to U.S. application Ser. No. 10/957,095 filed on Sep. 30, 2004, now U.S. Pat. No. 7,211,191.

FIELD OF THE INVENTION

The following invention relates generally to instrumentalities and methodologies in blood component separation. More specifically, the instant invention is directed to a method and apparatus for collecting a blood sample and subsequently separating the collected sample into constituent blood components for individual storage or use.

BACKGROUND OF THE INVENTION

Blood collection is always important, particularly in times of emergency (immediate use), but whole blood may only be stored for about 30 days before it is “outdated”. For long term storage, the ability to separate the whole blood into its major components (white blood cells, platelets, red blood cells and plasma) is of paramount importance because the long term storage condition for each component is different in terms of temperature and storage media. The most important component separations occurring after collection is the separation of red blood cells (RBC), white blood cells (WBC), platelets, and plasma from one another. Within the WBC it is sometimes important to separate the granulocytes from the lymphocytes and monocytes. After separation and extraction of particular components, a fraction of the blood may be returned to the patient.

It is possible to separate the various components of whole blood either under or after centrifugation, due to their differing densities. Some prior art methods, such as that in U.S. Pat. No. 4,120,448, utilize a chamber connected to a centrifuge. The centrifuged blood separates in the chamber, and a plurality of collection means are positioned at various locations in the chamber corresponding to the areas where each component congregates, which is density-dependent.

The present (prior art) technique for sequestering white blood cells from whole blood: requires skilled technicians, is labor intensive in that it requires 16 steps conducted over the span of one hour, and produces inconsistent results because of the requirements placed on the technician in the exercise of technique. Most significantly, however, the 16 step present technique is “open”; that is, the blood product is processed in a manner that does not maintain the sterility of the product because the need to obtain samples or add sedimenting agents or cryoprotectants at the various stages of production can not be accomplished with allowing the outside environment access to the interior, meaning potential contamination of the product:

The 16 steps are:

2. Add HES to collection bag (20% v/v).

3. Load collection bag into special centrifuge cup supports.

4. Centrifuge at 50 G for 13 min. to raise WBC from RBC (up to 6 units at one time).

5. Spike or sterile dock collection bag to expressor and processing bag set to scale.

6. Gently transfer collection bag to expressor and processing bag set to scale.

7. Express off WBC rich plasma and 10-15 ml of the top layer of RBC into processing bag—leaving excess RBC.

8. Remove collection bag with excess RBC.

9. Load processing bag set in special centrifuge cup supports.

10. Centrifuge processing bag set at 400 G for 10 min. (up to 6 units at one time).

11. Gently transfer processing bag to expressor.

12. Express off excess plasma leaving 20 ml WBC concentrate.

13. Remove excess plasma bag from processing set.

14. Add 5 ml cryoprotectant to WBC in processing bag at 4° C.

16. Tube seal and separate freezing bag from processing bag.

The following prior art reflects the state of the art of which applicant is aware and is included herewith to discharge applicant's acknowledged duty to disclose relevant prior art. It is stipulated, however, that none of these references teach singly nor render obvious when considered in any conceivable combination the nexus of the instant invention as disclosed in greater detail hereinafter and as particularly claimed.

The prior art references listed above but not specifically described teach other devices for blood processing and further catalog the prior art of which the applicant is aware. These references diverge even more starkly from the reference specifically distinguished above.

SUMMARY OF THE INVENTION

The present invention comprises a bag set that may be used to collect a whole blood sample from a source. Most significantly, the bag set defines a closed system in that once the blood is introduced, processing can occur outside a clean room or away from a sterile hood because access to any pathogens in the exterior environment is prevented. The bag set is then placed into a centrifuge for component separation. The whole blood processing bag, which may contain an anticoagulant such as CPD, ACD or CPD-A, contains at least one inlet and one outlet port connected to a plurality of component bags. The processing bag may optionally contain a sedimenting aid such as HES, but, unlike the prior art, such sedimenting aid is not required. Each component bag has a separate line leading from the whole blood processing bag, and each line can be clamped, tube-sealed and separated from the whole blood processing bag once a particular component bag has been filled.

In practice, the blood is collected and directed into an inlet port on the whole blood processing bag and the input line is clamped, sealed off, and separated from the whole blood processing bag. The whole blood processing bag, which is asymmetrically shaped, hangs in a bag set holder having a complementally shaped opening that closely contacts the bag at the bottom end, and an exterior of the bag set holder is adapted to fit in a conventional centrifuge cup or socket. The centrifuge is operated at varying G-forces to optimally separate the components. Once the components are separated by density in the whole blood processing bag, a servo motor is engaged to open a metering valve on the line leading from the processing bag to a bag that will contain the densest component. This allows the densest component to fill its particular storage bag, usually under centrifugation.

Applicant's process can be summarized in the following 7 or 8 steps which are performed over a span of 25 minutes, resulting in repeatable yields in excess of 90% of the lymphocytes and monocytes.

2. Spike or sterile dock collection bag to bag processing set and transfer blood to processing bag.

4. Load processing bag set onto auto expresser.

5. Centrifuge at an uninterrupted Run at two different speeds: 1,400 G for 20 min. to segregate WBC at RBC/plasma interface and 85 G for 5 min. to express the RBC to the RBC bag and WBC to freezing bag.

6. Tube seal and separate excess RBC and plasma bags from processing set.

7. Add 5 ml cryoprotectant to WBC in freezing bag at 4° C.

8. Tube seal and separate freezing bag from cryoprotectant line.

Complete collection of the first component is indicated preferably by an optical sensor that is present in the bag set holder device. The servo motor, directed by the sensor, automatically closes the metering valve on the line, terminating collection of that particular component. The servo motor then further engages the metering valve to allow collection of the next component through a second output line connecting the metering valve and the second storage bag. The process may sequentially continue until all desired components are collected in separate storage bags: red blood cells, white blood cells (lymphocytes and granulocytes), platelets, and plasma. If so desired, multiple components, such as the white blood cells and the platelets can be directed to the same storage bag. The sensor may be other than optical. For example, the sensor may monitor changes in electrical characteristics inherent in differing densities, such as capacitance, viewing the fluid as a dielectric. Commercially available markers (e.g. monoclonal antibodies, polarized particles, magnetic density, or fluorescence markers, etc.) can be introduced into the blood and monitored.

The bags receiving fluid components may also be supported for weighing both during centrifugation and when at rest. Accurate separation occurs.

Once collected, each storage bag may be sealed off and separated from the whole blood processing bag. Any necessary preservatives or additives may be introduced through the collection lines before processing or storing.

OBJECTS OF THE INVENTION

Accordingly, it is a primary object of the present invention to provide a new and novel device and method for separating the components of whole blood for subsequent storage or use.

It is a further object of the present invention to provide a device and method as characterized above in which separation may be accomplished entirely by machine during a single uninterrupted centrifugation run without the considerable handling between multiple centrifugation runs typically practiced in a blood bank with conventional means of separating blood components.

A further object is to precisely sequester red blood cells, plasma, platelets and white blood cells even separating within white blood cell populations.

It is a further object of the present invention to provide a device and method as characterized above in which the separation apparatus is self-contained to simplify the operation.

Viewed from a first vantage point, it is an object of the present invention to provide a device for sequestering components from whole blood, comprising, in combination: a bag set, said bag set including a first bag and plural other bags; a bag set holder, whereupon the first bag is contained within an interior portion of the bag set holder, and the plural other bags are located at an elevation lower than said first bag; and a centrifuge having at least two diametrically opposed receiving sockets, at least one socket dimensioned to receive the bag set holder.

Viewed from a second vantage point, it is an object of the present invention to provide an apparatus for use with a conventional centrifuge and a blood processing bag set, comprising, in combination: a first pocket having an unenclosed top portion, the first pocket dimensioned to receive a blood processing bag; means to support the blood processing bag in the first pocket, the support means located adjacent the unenclosed top portion of the first pocket; a movable bottom portion below the first pocket, the movable bottom portion having an open position and a closed position; a hinged portion located along a long axis of the first pocket, the hinged portion opening to allow access to the first pocket when the movable bottom portion is in the open position; and a second pocket, wherein access to the second pocket is only possible when the movable bottom portion is in the open position.

Viewed from a third vantage point, it is an object of the present invention to provide a method for separating components from whole blood, the steps including: preparing a blood processing bag set having a processing bag, at least one auxiliary bag, a sampling site adjacent the processing bag, and a sampling site adjacent each auxiliary bag; introducing whole blood into the processing bag; sampling the whole blood for later analysis; centrifuging the whole blood, wherein components are separated in the processing bag; directing each component into the at least one auxiliary bag of the blood processing bag set; removing a sample of each component for later analysis; and storing each component for later use.

Viewed from a fourth vantage point, it is an object of the present invention to provide a bag set, comprising, in combination: a first bag having an inlet and an outlet; plural auxiliary bags, each auxiliary bag having at least one port for admitting or expelling contents of the auxiliary bags; conduit means leading from the first bag to each auxiliary bag; valve means on the conduit means, the valve means adjustable to allow selective access between the first bag and the plural auxiliary bags.

These and other objects will be made manifest when considering the following detailed specification when taken in conjunction with the appended drawing figures.

DESCRIPTION OF PREFERRED EMBODIMENTS

Considering the drawings, wherein like reference numerals denote like parts throughout the various drawing figures, reference numeral10as shown inFIG. 3is directed to the bag set according to the present invention.

In its essence, the bag set10includes a whole blood processing bag2, a red blood cell (RBC) bag4having a hanger16, and a freezing bag6for the collection and storage of white blood cells. The processing bag2is supplied through an inlet line12, either through a phlebotomy needle8(FIG. 10) or by being spiked, or sterile docked, to another bag containing the anti-coagulated blood. The processing bag2has an asymmetric shape including a top edge11a, a short side edge11b, a long side edge11c, and a sloped bottom edge11dbetween the side edges such that the bottom portion tapers to an asymmetric point14, which leads to an outlet26.

Asymmetric processing bag allows concentration of a monocular cell fraction of a white cell population in a time frame that excludes 30-50% of the granulocyte white cells. Granulocytes have no role in the hematopoietic reconstitution and, thus their deletion results in a more purified selection of white cells for transplant.

Also, the asymmetric bag set allows this purification to take place without the need for a sedimenting agent—which is too viscous to sterilize through a filter—thus allowing the MNC to be concentrated in a “closed” sterile bag set as the DMSO can be made sterile by passage through a 0.2μ filter at the cryoprotectant inlet to the bag set.

The outlet26directs output from the processing bag2into a three-way metering valve20. The operating positions of the metering valve20are shown inFIGS. 14A-14C. Two supply lines24a,24blead from the metering valve20to the RBC bag4and the freezing bag6, respectively. The supply lines24a,24band the inlet line12may each be heat sealed and separated from the bag set10. All lines are equipped with line clamps22that may be closed to prevent fluid passage when desired. If other components are to be separated, the bag set10may include additional bags with a corresponding adjustment to the metering valve20to accommodate the additional bags.

Various supply lines may also be present in the bag set10. For example, the freezing bag supply line24bmay have an inlet16for the introduction of cryoprotectant into the system. Such inlets may be equipped with filters30(see, e.g.,FIG. 10), preferably 0.2μ filters, to, inter alia, prevent contamination from pathogens in the outside air and to allow venting of air from the freezing bag and tubing. An intermediate buffycoat bag40(FIG. 10) may be present on the freezing bag supply line24b. The buffycoat bag40collects a separate white cell fraction, which includes platelets and white cells and includes some small volume of plasma or red blood cells.FIGS. 10aand11ashow the bag set without the intermediate buffycoat bag40.

Initially, the processing bag2is either filled with an anticoagulant, such as CPD (citrate, phosphate, and dextrose) and blood is drawn through a phlebotomy needle into the bag, or the inlet line is spiked or sterile docked to another bag containing anticoagulated blood. The metering valve20begins in the closed position (FIG. 8A). All clamps22are closed with the exception of the clamp22on the inlet line12. Blood, preferably peripheral, placental umbilical cord blood, or bone marrow is obtained from a source through the phlebotomy needle8or other appropriate inlet, which feeds into the processing bag2through the inlet line12. The inlet line12is then clamped, heat sealed, and separated from the bag set10. Optionally, HES may be introduced into the RBC bag4through an optional inlet either before or after blood collection.

At this point, the bag set10is placed in a bag holder50, shown inFIGS. 1,2. The bag holder50is somewhat cylindrical, having a substantially elliptical shape, having two rounded ends connected by substantially straight sides. The main compartment70has an elongated oval shape dimensioned to receive the processing bag2. The main compartment70is accessed by sliding down a bottom portion162of the bag holder50(along arrow Z), then opening a cover72about a hinge71(along arrow X) present at one of the rounded ends of the bag holder50. The processing bag2is oriented in the bag holder50such that the hinged cover72closes over the edge11ccoinciding with the point14leading to the metering valve20. The metering valve20is received in an orifice74alocated on the major portion of the bag holder50. A complimental orifice74b, located on the hinged cover72, receives the protruding end of the metering valve20. The hinged cover72will only close when the bottom portion162is in the closed position. When the bottom portion is closed, a notch164in the bottom portion162registers with a retaining tab166present on the main body of the bag holder50.

Referring toFIG. 1, the bag holder50includes a bag hanger76having hooks60that engage the loops28on the processing bag2, maintaining the bag in position during the centrifuging process. The main compartment70of the bag holder50is shaped to receive the processing bag2, having a sidewall156that is complemental to the asymmetric shape of the processing bag2, which terminates in an outport160dimensioned to receive the asymmetric point14and the outlet26of the processing bag2. The sidewalls156cradle the processing bag2loosely around the middle and more tightly at the bottom (near the outlet26). Closer tolerance near the bottom of bag2is desired to minimize disturbing the contents of the bag after sedimentation. Thus, the top of compartment70mirrors the exterior elliptical shape but tapers down to the outport160while maintaining bag edges11b,11c,11din supporting relationship.

A notch78is present along one of the substantially straight sides of the bag holder50. The notch78receives the hanger16on the RBC bag4. The RBC bag4hangs along the outside of the bag holder50in a curved recess80leading to a lower support shelf83via transition81. The freezing bag6is cradled in a receptacle82located beneath the main compartment70of the bag holder50, accessed by sliding the bottom portion162down to open along arrow Z.FIGS. 4 and 5show the entire bag set10loaded in the bag set holder50before component separation occurs.FIG. 37shows a further iteration of a bag set showing schematically that the freezer bag is weighed during the separation process.FIG. 38shows the freezer bag has been encapsulated in a shell501which depends from platform503that supports, on its top side a control chip module57and on its bottom side the shell and freezer bag via a weighing load cell505. Shell501floats in an air space508, protected by “U” shaped bracket509.

The metering valve20is connected to a motor driver56in the bag holder50. The servo motor56is connected to a software-controlled control chip module57powered by a rechargeable battery B. Module57may require temperature compensation due to heat generation during centrifugation. A port P is provided to utilize a battery charger C (FIG. 35). The servo motor56controls the operation of the metering valve20while the bag set10is mounted in the bag holder50. One or more optical sensors58trigger the proper time for the servo motor56to close the metering valve20after each fraction is harvested. The sensor may be present at the position shown inFIG. 1or lower, closer to the outport160(FIG. 8C) adjacent the asymmetric point14of the processing bag2. Sensors58, for example may monitor all branches around valve20and the inlets of bags4and6. The sensor58shown is optical but can be based on density, weight, infrared, radioactivity, fluorescence, color, magnetism, ultrasonics, capacitance, wherein the characteristic measured may be an additive.

The bag holder50, when closed, is adapted to fit into a centrifuge cup66dimensioned to reside within a conventional centrifuge100. Preferably, at least two bag set holders50are placed in diametrically opposed centrifuge cups66, as shown inFIG. 6, for balance. A bag set10in the centrifuge cup66may be subjected to more than one G-force in order to achieve the optimum stratification of components (FIGS. 8A-8C). The servo motor56then operates the metering valve20to open and allow access to supply line24afor the harvest of red blood cells, at an optimum G-force, into bag4. The servo motor56closes the metering valve20when the optical sensor58indicates that the red blood cells are harvested (FIGS. 8A,8B). The optical sensor58senses the boundary between the white cell fraction and the plasma fraction.

The next fraction, which includes white cells and/or platelets, is then harvested from the processing bag2; the servo motor56opens the metering valve20to allow access to supply line24b(FIG. 8C) leading to bag6for the next harvest. As shown inFIG. 9, during the harvest (WBC) into the freezing bag6, air in the supply line adds to air already in the freezing bag6, producing an air bubble70, which is useful to assist the proper mixing of the WBC and/or platelets with the cryoprotectant. The servo motor56then closes the metering valve20, as shown inFIG. 8A, and the centrifuge100is allowed to stop.FIG. 9shows the bag set10in the bag set holder50after component separation has taken place.

The buffycoat bag40, if present, preferably has a 25 ml capacity. 20 ml of buffycoat is introduced into the buffycoat bag40, and 5 ml of DMSO solution is subsequently introduced. The buffycoat bag is placed between two cold strata and rotating or kneading of the buffycoat bag40in order to mix the cryoprotectant and WBC solution takes place.

The bag holder50is removed from the centrifuge cup66and opened, and the bag set10is removed, with the servo motor56disconnected from the metering valve20. Each supply line24a,24bis clamped, heat sealed, and removed from the processing bag2. Any additional bags may be similarly removed.

After the supply line24bconnected to the freezing bag6is disconnected, a cryoprotectant may be introduced into the collected component in the freezing bag6through an inlet. The air bubble70in the freezing bag6allows the cryoprotectant to be thoroughly mixed with the collected component. After mixing, the air bubble70is expelled, perhaps through a filter-protected cryoprotectant inlet16(FIG. 10). The component is then prepared for storage by heat-sealing the tubing and removing the bag6downstream of the cryoprotectant inlet16.

Preferably, each line (the inlet line12and the supply lines24a,24b) is oriented to allow access to a sampling site (e.g., site18) near the collection or storage bags. Thus, a sample of the blood or fluid in the line may be taken without disturbing the bulk of the collected component.

FIG. 13depicts the separation of whole blood components as a function of time. Under centrifugation, each fraction stratifies in the processing bag2as a function of its density. The overlapping areas175(FIG. 13) indicate the area in the separation along each strata line in the processing bag2. As centrifugation continues, the boundary of each fraction becomes more clearly defined; thus, the area175(FIG. 13) decreases and each fraction is more completely harvested. Thus, the centrifugation strategy combines separation by density, the time involved for stratification, which differs with the exterior surface area and density of the various cells, centrifugal force, and boundary layer clarity. Decisions on harvesting will vary based on these tradeoffs as a function of the constituent of greatest value and its desired purity.

Preferably, the stratification centrifugation occurs at an excess of 1000 Gs, preferably 1400 Gs, for approximately 20 minutes. The transfer centrifugation step occurs at less than 100 Gs, preferably 78 Gs, and stops subject to output from the optical sensor58. The right hand side ofFIG. 36shows the white cell bag (Freezer bag6) topped off in increments by throttling the valve20on and off in order to extract the WBC population.

It is appreciated that while the instant invention is preferably used in the separation of blood components, the separation techniques and apparatus are suitable for separation of other fluids. The software programmed into the control chip module may cause the servo motor to open and close the valve many times, thereby throttling the valve during strata delivery. Also by varying time increments during a harvest procedure, precise cut-offs between the cell components can be achieved in order to reduce the mixing between cell types that may occur as a result of the “toroidal” (Coriolis) effect during removal of the blood component from processing bag2and may be modified for the separation of other fluids or to compensate for various hardware conditions, such as uneven centrifuge loading.

Yet another embodiment of the bag set210is shown inFIG. 15. In its essence, the bag set210includes a whole blood processing bag202, a red blood cell (RBC) bag204, and a freezing bag206. The processing bag202is supplied through an inlet line212that terminates in a spike208. The processing bag202has an asymmetric shape including a top edge211a, a short side edge211b, a long side edge211c, and a sloped bottom edge211dbetween the side edges such that the bottom portion tapers to an asymmetric point214, which leads to an outlet226. The outlet226directs output from the processing bag202into a stopcock valve220. Two supply lines224a,224blead from the stopcock valve220to the RBC bag204and the freezing bag206, respectively. The supply lines224a,224band the inlet line212may each be heat sealed and separated from the bag set210. All lines are equipped with line clamps222that may be closed to prevent fluid passage when desired. If other components are to be separated, the bag set210may include additional bags with a corresponding adjustment to the stopcock valve220to accommodate the additional bags.

Initially, the blood of interest is collected in a collection bag200or similar container. The spike208is inserted into the collection bag200, and the blood is drained from the collection bag200into the processing bag202through the inlet line212(FIGS. 16,17). The inlet line212preferably has a clot filter230, through which the blood passes before it reaches the processing bag202. After the blood is transferred, the inlet line212is heat sealed and the collection bag200and clot filter230are removed (FIG. 18).

The inlet line212also preferably has a sampling port232, a sampling pillow234, and an access port236(FIG. 19). After the collection bag200and clot filter230are moved from the inlet line212, the sampling pillow234is squeezed and released to fill the sampling pillow with blood. The inlet line212is then heat sealed and the sampling pillow234is removed, along with the sampling port232(FIG. 20). The blood in the sampling pillow234may then be accessed through the sampling port232for separate assay.

Unlike the prior art where a sedimentation agent is required, a sedimenting agent, such as hydroxyethyl starch (HES) may optionally be added to the processing bag202through the access port236on the inlet line212using syringe means236aor similar delivery means, and the processing bag202is manipulated to thoroughly mix the agent with the blood (FIG. 21). The bag set210is then placed into the bag holder50and used with a centrifuge, as detailed hereinabove, to separate the cells therewithin (FIG. 22). The separated red blood cells are transferred into the RBC bag204and the WBC fraction is transferred to the freezing bag206during this operation. The bag set210is then removed from the bag holder50(FIG. 23). Supply line224ais then heat sealed and the RBC bag204is removed (FIG. 24). The contents of the RBC bag are accessed through a sample port238.

Referring toFIG. 25, supply line224bis preferentially equipped with a first junction260connecting an auxiliary inlet line240terminating in an auxiliary port242. A second junction262is present on the auxiliary inlet line240itself to connect a branch line244that terminates in a bulb246. The branch line244also contains a sampling pigtail248and a sampling port250. After removal of the RBC bag204, the bulb246on the branch line244is squeezed to direct any residual plasma remaining in the supply line224binto the freezing bag206. Clamp222on branch line244is then closed. The contents of the freezing bag206are then mixed, preferably by holding the freezing bag206at a 45° angle and slowly squeezing the small compartment206aof the freezer bag206a total of ten times at one squeeze per second.

The clamp222on the branch line244is then opened, and the bulb246is squeezed and released to fill the sampling pigtail248with the contents of the freezer bag206(FIG. 26). The branch line244is heat sealed and removed from the bag set210(FIG. 27). The contents of the sampling pigtail248are accessed through the sampling port250for separate assay.

The freezing bag206is placed on its side and sandwiched between two ice packs252(FIG. 28). DMSO is introduced into the freezing bag206through the auxiliary port242which has a sterile filter242a(i.e. less than or equal to 0.2 microns) on the auxiliary inlet line240. An orbital mixer254is used with the sandwiched freezer bag206to thoroughly mix the contents of the freezer bag206. The sandwiched freezer bag206is then placed in stationary holder256(FIG. 29). A syringe258is inserted into the auxiliary inlet242and used to draw out any residual DMSO and trapped air in the supply line224band the auxiliary inlet line240. The buffy coat/DMSO from the freezing bag206is drawn out by the syringe258until it reaches the second junction262from the supply line224b. The freezing bag206is then removed from the bag set210by heat sealing the supply line224b(FIG. 30).

A portion of the supply line224bafter the first junction260remains attached to the freezing bag206. This portion of the supply line224bis heat sealed to form three separate samples275a,275b,275c(still connected to the freezing bag206), and the area separating the small compartment206aof the freezer bag206is heat sealed to separate it from the rest of the freezer bag206(FIG. 31). The final product is then frozen for storage.

The stopcock valve220is turned to allow plasma in the processing bag202to contact the buffy coat in the supply line224bnear the first and second junctions260,262(FIG. 32). A sample of the plasma diluted buffy coat is drawn into the syringe258for bacterial sampling, and the syringe258is removed from the auxiliary port242. The supply line224bcontaining the auxiliary line240and the first and second junctions260,262is then disconnected from the processing bag202and is discarded (FIG. 33). Samples of the plasma in the processing bag202may be removed by using the access port236(FIG. 34).

Moreover, having thus described the invention, it should be apparent that numerous structural modifications and adaptations may be resorted to without departing from the scope and fair meaning of the instant invention as set forth hereinabove and as described hereinbelow by the claims.