Cell culture apparatus

A cell culture apparatus including cell culture units U and a measurement part (5). The measurement part (5) is provided with an imaging part (51) and is disposed so as to be capable of moving up and down between a back panel part (21) of a casing (2) and a culture container accommodating shelf (32). When the measurement unit (5) reaches a measurement position of each cell culture unit U which is a measurement subject, the culture container accommodating shelf (32) slides toward the measurement unit (5) so that portions to be measured of culture containers U20, U30 in each cell culture unit U come in an imaging range of the imaging part (51).

CROSS REFERENCE TO RELATED APPLICATIONS

This Application is a U.S. National Stage of Application No. PCT/JP2015/004591, filed on Sep. 9, 2015. Priority under 35 U.S.C.§ 119(a) and 35 U.S.C.§ 365(b) is claimed from Japanese Patent Application No. 2014-189273 filed on Sep. 17, 2014, Japanese Patent Application No. 2014-189274 filed on Sep. 17, 2014, Japanese Patent Application No. 2014-189275 filed on Sep. 17, 2014 and Japanese Patent Application No. 2014-189276 filed on Sep. 17, 2014, the disclosures of which are incorporated herein by reference in their entirety.

FIELD

The present invention relates to a cell culture apparatus.

BACKGROUND

In recent years, it has been required that cells, tissues or the like are efficiently cultured in large amounts under an artificial environment in fields of production of medicines, gene therapy, regenerative therapy, immunotherapy and the like.

In such cell culture, a medium container to supply a medium to a culture container and the culture container for use in cell culture are connected by a tube to constitute a cell culture unit, and the cell culture is performed in a closed system by use of this cell culture unit, thereby avoiding a risk of contamination to proliferate the cells.

In this cell culture unit, the medium is generally required to be refrigerated, and hence the medium container is stored in a cold insulation chamber. Furthermore, in the cell culture, it is necessary to control a temperature, air environment and the like, and hence the culture container is stored in an incubator. Then, the medium is transferred from the medium container to the culture container via the tube.

Alternatively, for example, in Patent Literature 1, a cell culture apparatus is disclosed in which such culture units are stored in a multi-stage state and cell culture is performed.

RELATED ART DOCUMENT

Patent Literature

SUMMARY

Technical Problem

However, in a culture apparatus described in Patent Literature 1, for the purpose of observing the number and others of cells proliferated in a culture container (a culture bag), the cells in the culture container are guided to another container (a cell inoculation cassette or a dummy cassette) and imaged by using a CCD camera. In addition, all culture units have a similar constitution, and a complicated apparatus constitution is taken.

The present invention has been developed in view of the above circumstances, and an object thereof is to provide a cell culture apparatus in which a plurality of cell culture units each comprising culture containers to which a medium is supplied together with cells to perform cell culture and a medium container to supply the medium to the culture containers are stored in a multi-stage state, and when cell culture is individually performed in each of the cell culture units, a progressive state of the cell culture in all the cell culture units can be measured by a single measurement unit.

Solution to Problem

To achieve the above object, the present invention is directed to a cell culture apparatus in which there are stored, in a multi-stage state in a casing, a plurality of cell culture units each comprising at least a culture container to which a medium is supplied together with cells to perform cell culture and a medium container to supply the medium to the culture container, the culture container and the medium container being connected by a transfer tube, thereby individually performing the cell culture in each of the cell culture units, wherein a support frame in which a plurality of slide rails are arranged in a multi-stage state along a height direction is disposed in the casing, a culture container accommodating shelf in which the culture containers are accommodated is slidably attached to each of the slide rails, a measurement unit comprising an imaging part is disposed so as to be movable up and down between a back panel part of the casing and the culture container accommodating shelf, and when the measurement unit reaches a measurement position of each cell culture unit that is a measurement subject, the culture container accommodating shelf which accommodates the culture containers provided in the cell culture unit moves toward the measurement unit so that portions to be measured of the culture containers provided in the cell culture unit come in an imaging range of the imaging part.

According to such a constitution, the plurality of cell culture units each comprising the culture containers and the medium container are stored in the multi-stage state, and when the cell culture is individually performed in each of the cell culture units, a progressive state of the cell culture in the cell culture unit of the measurement subject can be measured by a single measurement unit in each of all the cell culture units without interfering with an adjacent cell culture unit.

Furthermore, in a culture apparatus described in Patent Literature 1 mentioned above, a medium container (a medium bag) and a culture container (a culture bag) are stored in the same thermostatic tank, so cell culture cannot be performed under a temperature environment suitable for each of these containers.

In view of the above circumstances, the present invention can have the following constitution to provide a cell culture apparatus in which a plurality of cell culture units each comprising culture containers to which a medium is supplied together with cells to perform cell culture and a medium container to supply the medium to the culture containers are stored in a multi-stage state, and when the cell culture is individually performed in each of the cell culture units, the cell culture can efficiently be performed under a temperature environment suitable for each of the culture containers and the medium container.

That is, the cell culture apparatus according to the present invention may have a constitution comprising an incubating tank in which the culture containers are stored and a cold insulation tank in which the medium container is stored, wherein in each of the incubating tank and the cold insulation tank, there are disposed an incubating tank side support frame and a cold insulation tank side support frame in each of which a plurality of slide rails are arranged in a multi-stage state along a height direction, the culture container accommodating shelf is slidably attached to each of the slide rails arranged in the multi-stage state in the incubating tank side support frame, and a medium container accommodating shelf to accommodate the medium container is slidably attached to each of the slide rails arranged in the multi-stage state in the cold insulation tank side support frame, and the culture container accommodating shelf and the medium container accommodating shelf form a pair and are disposed separately in the incubating tank and the cold insulation tank.

According to such a constitution, the plurality of cell culture units each comprising the culture containers and the medium container are stored in a multi-stage state, and when the cell culture is individually performed in each of the cell culture units, the cell culture can efficiently be performed under the temperature environment suitable for each of the culture containers and the medium container.

Alternatively, the medium container and the culture containers are connected by the tube to constitute the cell culture unit, and when the cell culture is performed in a closed system, bubbles might enter into the culture containers in a process of the cell culture, for example, at a time of supplying the medium from the medium container to the culture containers. When the bubbles enter into the culture containers, it is feared that a culture environment in the culture container does not become uniform, and an efficiency of the cell culture deteriorates. It is also considered that, when a content is taken out from the culture container after the cell culture is ended, the liquid delivery is disturbed by the bubbles, but in the culture apparatus described in Patent Literature 1 mentioned above, these problems are not taken into consideration at all.

For example, it is considered that the bubbles flow into the medium container or the like which forms the closed system together with the culture container, for the purpose of removing the bubbles which enter into the culture container while maintaining the closed system. However, when a timing to end an operation of removing the bubbles is wrong, the bubbles might remain in the culture containers, and the contents of the culture containers might flow into the medium container or the like.

In view of the above circumstances, the present invention can have the following constitution to provide a cell culture apparatus in which a cell culture unit comprising culture containers to which a medium is supplied together with cells to perform cell culture and a medium container to supply the medium to the culture containers is stored, and when the cell culture is performed in this cell culture unit and when an operation of removing bubbles which enter into the culture containers in a process of the cell culture is performed, a timing to end the operation can be judged.

That is, the cell culture apparatus according to the present invention may have a constitution in which the culture container accommodating shelf or the medium container accommodating shelf comprises weight detecting means, and in removing bubbles which enter into the culture containers in the process of the cell culture, the bubbles are transferred to the medium container through the transfer tube, and the weight detecting means provided in the culture container accommodating shelf or the medium container accommodating shelf detects a weight change of the culture container or the medium container to end a transfer operation of the bubbles.

According to such a constitution, the cell culture unit comprising the culture containers and the medium container is stored, and when the cell culture is performed in this cell culture unit and when an operation of removing the bubbles which enter into the culture containers in the process of the cell culture is performed, the timing to end the operation can be judged and the operation can automatically or manually be ended.

Furthermore, when the contents of the culture containers are taken outside in addition to removing the bubbles which enter into the culture containers in the process of the cell culture, it is required that the contents are inhibited from remaining, but in the above-mentioned culture apparatus described in Patent Literature 1, these problems are not taken into consideration at all.

In view of the above circumstances, the present invention can have the following constitution to provide a cell culture apparatus in which there is stored a cell culture unit comprising culture containers to which a medium is supplied together with cells to perform cell culture and a medium container to supply the medium to the culture containers, and when the cell culture is performed in this cell culture unit, bubbles which enter into the culture containers in a process of the cell culture are easily removed, and when contents of the culture containers are taken outside, the contents can be inhibited from remaining in the culture containers.

That is, the cell culture apparatus according to the present invention may have a constitution in which the culture container accommodating shelf comprises tilting means for tilting the culture containers so that ports for liquid delivery provided in the culture containers are positioned at lower positions or upper positions.

According to such a constitution, the cell culture unit comprising the culture containers and the medium container is stored, and when the cell culture is performed in this cell culture unit, the bubbles which enter into the culture containers in the process of the cell culture can easily be removed, and when the contents of the culture containers are taken outside, the contents can be inhibited from remaining.

Advantageous Effects of Invention

According to the present invention, a plurality of cell culture units each comprising culture containers and a medium container are stored in a multi-stage state, and when cell culture is individually performed in each of the cell culture units, a progressive state of the cell culture in each cell culture unit that is a measurement subject can be measured by a single measurement unit in each of all the cell culture units without interfering with an adjacent cell culture unit.

DETAILED DESCRIPTION

Hereinafter, a preferable embodiment of a cell culture apparatus according to the present invention will be described with reference to the drawings.

In the present embodiment, a cell culture apparatus1is an apparatus to store a plurality of cell culture units U in a multi-stage state and individually perform cell culture in each of the cell culture units U.FIG. 1andFIG. 2show appearance schematic views of the cell culture apparatus1. In the cell culture apparatus1, an inner portion of a casing2has a double tank structure divided into an incubating tank3and a cold insulation tank4which can be adjusted at a predetermined temperature, by an insulation wall20. Further, the incubating tank3and the cold insulation tank4are provided with independently openable and closable front doors3aand4a, respectively.

For example, as shown inFIG. 3, a cell culture unit U to be stored in the cell culture apparatus1comprises a first culture container U20and a second culture container U30for use in cell culture, a medium container U10to supply a medium to the culture containers U20, U30, and a transfer tube U40and culture container connecting tubes U42to connect the containers U10, U20and U30.

The first culture container U20is a culture container into which cells are injected, to first perform cell culture, and is suitably usable as, for example, an activating culture container for use in the cell culture to activate the cells. In a case of activating floating cells such as lymphocytes by use of the first culture container U20as the activating culture container, a substance such as an anti-CD3 antibody to activate the cells is immobilized in a solid phase on a bottom surface in the first culture container U20.

The second culture container U30is a container into which, after the cell culture in the first culture container, a cell suspension in the first culture container is transferred, to perform the cell culture, and the second culture container is suitably usable as, for example, a proliferating culture container for use in the cell culture to proliferate the cells.

In this way, the first culture container U20is used as the activating culture container, and the second culture container U30is used as the proliferating culture container, thereby making it possible to suitably proliferate floating cells such as the lymphocytes.

In the present embodiment, these containers are connected to the transfer tube U40in the order of the medium container U10, the second culture container U30and the first culture container U20. It is to be noted that in a case where the containers are seen from a back side, the containers are connected to the transfer tube U40in the order of the first culture container U20, the second culture container U30and the medium container U10, and these orders are the same.

The medium container U10comprises a port U11for liquid delivery (including inward delivery and outward delivery), and is connected to the transfer tube U40through the port U11.

The first culture container U20comprises a port U21for liquid delivery, and is connected to the transfer tube U40through the port U21.

As shown inFIG. 3, it is preferable that the port U21is disposed on the side of the transfer tube U40in a peripheral edge portion of the first culture container U20and most apart from the second culture container U30. Consequently, in delivering a content out from the first culture container U20, i.e., in transferring the cell suspension from the first culture container U20to the second culture container U30, the first culture container U20is tilted so as to dispose the port U21downwardly, thereby making it possible to inhibit the cell suspension from remaining in the first culture container U20.

Furthermore, in removing bubbles which enter into the first culture container U20in a process of the cell culture, for example, at a time of supplying the medium from the medium container U10to the first culture container U20, the first culture container U20is tilted to dispose the port U21upwardly, whereby the bubbles which enter into the first culture container U20can be collected in the vicinity of the port U21and can easily be removed from the port U21.

The second culture container U30comprises a port U31and a port U32for liquid delivery, one side end portions of two culture container connecting tubes U42are connected to these ports respectively, and the other side end portions are connected to the transfer tube U40. A pump U41is attached to a portion between connected portions of the culture container connecting tubes U42in the transfer tube U40.

That is, the cell culture unit U comprises only one pump, and has a constitution in which the second culture container U30is connected to the transfer tube U40on both sides of the pump U41by the culture container connecting tubes U42to connect the second culture container U30and the transfer tube U40.

As shown inFIG. 3, it is preferable that the port U31is disposed on the side of the transfer tube U40in a peripheral edge portion of the second culture container U30and most apart from the medium container U10. Consequently, in delivering the content out from the second culture container U30, i.e., in transferring and collecting the cell suspension from the second culture container U30to the emptied medium container U10, the second culture container U30is tilted so as to dispose the port U31downwardly, thereby making it possible to inhibit the cell suspension from remaining in the second culture container U30.

Furthermore, in removing bubbles which enter into the second culture container U30in a process of the cell culture, for example, at a time of transferring the cell suspension from the first culture container U20to the second culture container U30or supplying the medium from the medium container U10to the second culture container U30, the second culture container U30is tilted to dispose the port U31upwardly, whereby the bubbles which enter into the second culture container U30can be collected in the vicinity of the port U31and can easily be removed from the port U31.

There is not any special restriction on the number of the ports in each of these containers, and the number of the ports can be adjusted to be optional. In an example ofFIG. 3, the first culture container U20comprises a port U22for cell injection in addition to the port U21. Furthermore, the second culture container U30comprises a preliminary port U33to inject a proliferation factor or the like and a sampling port U34in addition to the ports U31and U32.

When the liquid delivery is not performed between the containers, a flow channel from each container is closed with a closing member U44such as a clip. That is, in the example ofFIG. 3, the flow channel between each container and the pump U41is stopped with the clip. Further, in performing the liquid delivery between the containers, the closing with the closing member U44for the two containers between which the liquid delivery is performed is only released to open the flow channel, and the liquid delivery by the pump U41is performed.

To perform the cell culture in the cell culture unit U constituted in this way, the cells are first injected into the first culture container U20from the port U22.

Next, the pump U41is operated, and by the pump U41, the medium is transferred from the medium container U10to the first culture container U20as shown inFIG. 3(an arrow (1) in the same figure). Then, in the first culture container U20, the cell culture is performed for a predetermined period.

After the cell culture in the first culture container U20, the cell suspension is transferred from the first culture container U20to the second culture container U30by the pump U41(an arrow (2) in the same figure), and next the medium is transferred from the medium container U10to the second culture container U30(an arrow (3) in the same figure).

At this time, the transfer of the cell suspension from the first culture container U20to the second culture container U30is performed via the culture container connecting tube U42connected to the transfer tube U40on a medium container U10side of the pump U41.

Furthermore, the transfer of the medium from the medium container U10to the second culture container U30is performed via the culture container connecting tube U42connected to the transfer tube U40on a first culture container U20side of the pump U41.

According to the cell culture unit U, it is possible to appropriately perform, by use of only one pump U41, transfers of three systems, i.e., the transfer of the medium from the medium container U10to the first culture container U20, the transfer of the cell suspension from the first culture container U20to the second culture container U30and the transfer of the medium from the medium container U10to the second culture container U30.

In general, the medium container U10having gas barrier properties to oxygen and carbon dioxide is used so that a pH of the stored medium does not noticeably change in a culture period. Because, in order to avoid that a high-concentration carbonic acid gas included in the medium falls out in the air and that the concentration of the carbonic acid gas in the medium decreases to increase the pH, it is desirable to minimize leaking of carbon dioxide out from the medium container U10as much as possible, and it is also desirable to prevent oxidation of the medium.

The first culture container U20and the second culture container U30are formed into a bag shape by use of a soft packaging member as a material, and partially or entirely have transparency so that the contents can be confirmed. Furthermore, these culture containers are required to have a gas permeability (an oxygen and carbon dioxide permeability) required for the cell culture, and are preferably used at 37° C. under a culture environment where a concentration of carbon dioxide is 5%. Furthermore, it is preferable that the culture containers have low cytotoxicity, low elution, and radiation sterilization suitability to achieve a high cell proliferating efficiency.

As a material of the culture container which satisfies these conditions, a polyethylene-based resin is preferable. Examples of this polyethylene-based resin include polyethylene, a copolymer of ethylene and α-olefin, a copolymer of ethylene and vinyl acetate, and ionomers in which ethylene, an acrylic acid or methacrylic acid copolymer and metal ions are used. Alternatively, polyolefin, a styrene-based elastomer, a polyester-based thermoplastic elastomer, a silicone-based thermoplastic elastomer, a silicone resin or the like is usable.

There is not any special restriction on a shape of each of the medium container U10, the first culture container U20and the second culture container U30and a shape of a containing section of the container, and in the example ofFIG. 3, the container is formed into a rectangular shape. The container can be manufactured by sealing four sides of the container with a heat seal, or can be manufactured as an integrally formed bag by blow forming.

It is preferable that each port is disposed on the transfer tube U40side, and it is also preferable that the containing section gradually narrows toward an attaching portion of the tube.

A material of the transfer tube U40and the culture container connecting tubes U42may suitably be selected in accordance with a use environment, but is preferably excellent in gas permeability. For example, there is usable a silicone rubber, a soft vinyl chloride resin, a polybutadiene resin, an ethylene-vinyl acetate copolymer, a chlorinated polyethylene resin, a polyurethane-based thermoplastic elastomer, a polyester-based thermoplastic elastomer, a silicone-based thermoplastic elastomer, a styrene-based elastomer, or the like. Furthermore, as the styrene-based elastomer, there is usable, for example, SBS (styrene-butadiene-styrene), SIS (styrene-isoprene-styrene), SEBS (styrene-ethylene-butylene-styrene), SEPS (styrene-ethylene-propylene-styrene), or the like.

InFIG. 3, for example, a silicone tube is used in a portion of the transfer tube U40to which the pump U41is attached, and a soft vinyl chloride resin tube is suitably usable in another portion thereof.

Furthermore, connection between the tubes in the transfer tube U40can be performed with a luer connector or another coupling means. Furthermore, in the transfer tube U40, flow channel opening/closing means such as a pinch valve, a two-way stopcock or a three-way stopcock may be disposed.

There is not any special restriction on a type of pump U41, but a tube roller pump such as a peristaltic pump (R) is suitably usable because a closed system in the cell culture unit U is easy to be realized.

In the present embodiment, the cell culture unit U is stored in the cell culture apparatus1, but the first culture container U20and the second culture container U30are stored in the incubating tank3in which a temperature is adjusted to be maintained at a temperature (e.g., 37° C.) suitable for the cell culture, and the medium container U10is stored in the cold insulation tank4in which a temperature is adjusted to be maintained at a temperature (e.g., 4° C.) suitable for the storage of the medium.

As shown inFIG. 4, in the incubating tank3in which the first culture container U20and the second culture container U30are stored, an incubating tank side support frame30is disposed in which a plurality of slide rails31are arranged in a multi-stage state along a height direction. To each of the slide rails31arranged in the incubating tank side support frame30, a culture container accommodating shelf32to accommodate the first culture container U20and the second culture container U30is slidably attached to be pulled out to a proximal side. Consequently, when the first culture container U20and the second culture container U30are mounted on or removed from the culture container accommodating shelf32, the culture container accommodating shelf32is pulled out to the proximal side, thereby making it possible to easily perform such an operation.

It is to be noted that in an example shown inFIG. 4, the pump U41and the closing member U44constituting the cell culture unit U are attached to a proximal side of the culture container accommodating shelf32, but illustration of the transfer tube U40and others is omitted.

Furthermore, similarly in the cold insulation tank4in which the medium container U10is stored, a cold insulation tank support frame40is disposed in which a plurality of slide rails41are arranged in a multi-stage state along a height direction. Then, to each of the slide rails41arranged in the cold insulation tank side support frame40, a medium container accommodating shelf42to accommodate the medium container U10is slidably attached to be pulled out to the proximal side. In consequence, when the medium container U10is mounted on or removed from the medium container accommodating shelf42, the medium container accommodating shelf42is pulled out to the proximal side, thereby making it possible to easily perform such an operation (seeFIG. 5).

It is to be noted thatFIG. 5is a planar view showing a state where the medium container accommodating shelf42is pulled out to the proximal side, the closing members U44constituting the cell culture unit U are attached to the proximal side of the medium container accommodating shelf42, but illustration of the port U11and the transfer tube U40disposed in the medium container U10is omitted.

Consequently, in the present embodiment, the culture container accommodating shelf32and the medium container accommodating shelf42form a pair and are disposed separately in the incubating tank3and the cold insulation tank4which are adjusted at a predetermined temperature, the first culture container U20and the second culture container U30are accommodated in the culture container accommodating shelf32, and the medium container U10is accommodated in the medium container accommodating shelf42, whereby the plurality of cell culture units U can be stored in the multi-stage state. In consequence, according to the cell culture apparatus1of the present embodiment, under a temperature environment suitable for each of the first culture container U20, the second culture container U30and the medium container U10, the plurality of cell culture units U are independently stored in the multi-stage state, and the cell culture can individually be performed in each of the cell culture units U.

It is to be noted thatFIG. 2shows a state where the culture container accommodating shelf32and the medium container accommodating shelf42form the pair and are pulled out to the proximal side. However, for the convenience in preparing the drawing, illustration of the cell culture units U to be stored in theses shelves, auxiliary devices and others is omitted.

Furthermore, in the present embodiment, the incubating tank side support frame30is disposed to be capable of freely being inserted into and taken out from the incubating tank3so that the culture container accommodating shelves32and the other auxiliary devices attached to the incubating tank side support frame30can be taken together with the incubating tank side support frame30out from the incubating tank3. Similarly, the cold insulation tank side support frame40is also disposed to be capable of freely being inserted into and taken out from the cold insulation tank4, and hence the medium container accommodating shelves42and the other auxiliary devices attached to the cold insulation tank side support frame40can be taken together with the cold insulation tank side support frame40out from the cold insulation tank4. In consequence, the devices disposed in the casing2of the cell culture apparatus1can be taken out from and disposed in the respective tanks, together with the support frames30and40.

According to such a constitution of the cell culture apparatus1, for example, when maintenance or the like of the cell culture apparatus1is performed, the devices disposed in the casing2can be taken outside together with the support frames30and40, and an operation space on the side of the casing2can be acquired, so that an operability in performing the maintenance or the like of the cell culture apparatus1can improve. Furthermore, for example, at a time of generation of an abnormality in the cell culture apparatus1, the cell culture units U are not taken out one by one, but all the cell culture units U are taken out together with the support frames30and40, thereby making it possible to cope with the problem in a short time.

The support frames30and40are taken out from the casing2, and then these frames may be mounted on a cart to move, but the support frames30and40may be provided with moving means such as casters so that these frames are easily movable.

Furthermore, in the present embodiment, in the cell culture apparatus1in which the plurality of cell culture units U are stored in the cell culture apparatus1and the cell culture is performed in each of the cell culture units U, a measurement unit5to measure a progressive state of the cell culture in the first culture container U20and the second culture container U30provided in each of the cell culture units U is interposed between a back panel part21of the casing2of the cell culture apparatus1and the culture container accommodating shelf32(seeFIG. 4).

The measurement unit5comprises an imaging part51which images portions to be measured of the culture containers U20, U30, and there is not any special restriction on its specific constitution as long as the progressive state of the cell culture can be measured, for example, by analyzing imaging data and calculating the number of the cells, a cell density or the like. In the example shown in the drawing, the measurement unit5comprises the imaging part51having a CCD camera and an objective lens, and an illuminating part52disposed to face the imaging part51, and portions to be imaged of the culture containers U20, U30can be imaged from the downside, while illuminating the portions to be imaged of the culture containers U20, U30with illuminating light from the upside.

Although not especially shown in the drawing, in a case where the first culture container U20and the second culture container U30are formed into the bag shape by use of the soft packaging member as the material as in the present embodiment, the measurement unit5may comprise a pressing member which presses the portions to be imaged so that inner portions of the culture containers U20, U30can clearly be imaged in a state where bends of the portions are eliminated, when the portions to be imaged of the culture containers U20, U30are imaged.

The measurement unit5is disposed so as to be movable up and down in the casing2so that the progressive state of the cell culture can be measured in each of all the cell culture units U stored in the cell culture apparatus1. For example, in a support pillar50attached to the incubating tank side support frame30, the measurement unit is disposed so as to be movable up and down by a power source such as a motor.

Furthermore, in the present embodiment, the cell culture unit U comprises the first culture container U20and the second culture container U30, but the measurement unit5can be adjusted to be also movable in a horizontal direction as shown by arrows inFIG. 8so that the progressive state of the cell culture in each of the culture containers U20, U30can be measured.

In an example shown inFIG. 8, the measurement unit5is held by guide members50aand50bso as to be movable in the horizontal direction, and the guide members50aand50bare vertically movably attached to the support pillar50, but a specific mechanism is not limited to this example.

Alternatively, in the present embodiment, the culture container accommodating shelf32which accommodates the first culture container U20and the second culture container U30can not only be pulled out to the proximal side but also be slid toward a distal side. Consequently, when the measurement unit5moves in the casing2to reach a measurement position of the cell culture unit U which is the measurement subject, the culture container accommodating shelf32which accommodates the first culture container U20and the second culture container U30of the cell culture unit U moves toward the measurement unit5(seeFIG. 4andFIG. 8). Successively, when the portions to be measured of the first culture container U20and the second culture container U30accommodated in the culture container accommodating shelf32come in an imaging range of the imaging part51provided in the measurement unit5, the measurement of the progressive state of the cell culture by the measurement unit5is started.

In consequence, the progressive state of the cell culture in the cell culture unit of the measurement subject can be measured by a single measurement unit5in each of all the cell culture units U without interfering with an adjacent cell culture unit U.

Here,FIG. 6toFIG. 8are planar views showing states where the culture container accommodating shelf32is slid to move. When the culture container accommodating shelf32is pulled out to the proximal side from the state shown inFIG. 6, it becomes the state shown inFIG. 7. This state corresponds to a position of the culture container accommodating shelf32shown in a lowermost stage inFIG. 4. Successively, when the culture container accommodating shelf moves from the state shown inFIG. 6toward the measurement unit5, it becomes the state shown inFIG. 8. This state corresponds to a position of the culture container accommodating shelf32shown in the fourth stage from the bottom inFIG. 4.

It is to be noted that in an example shown inFIG. 6toFIG. 8, the pump U41and the closing members U44which constitute the cell culture unit U are attached to the proximal side of the culture container accommodating shelf32, but there is omitted illustration of the ports U21and U22disposed in the first culture container U20, the ports U31, U32, U33and U34disposed in the second culture container U30, the transfer tube U40and the culture container connecting tubes U42.

Furthermore, as described above, in transferring the content of the first culture container U20or the second culture container U30from its port U21or U31, the culture containers U20, U30are tilted so as to dispose the ports U21, U31downwardly, whereby the content can be inhibited from remaining. Additionally, in removing the bubbles which enter into the culture containers U20, U30, the culture containers U20, U30are tilted to dispose the ports U21, U31upwardly, whereby the bubbles which enter into the culture containers U20, U30can be collected in the vicinities of the ports U21, U31and can easily be removed.

In the present embodiment, for the purpose of enabling this operation, the culture container accommodating shelf32comprises a swing plate33as tilting means for tilting the first culture container U20and the second culture container U30so that the ports U21, U31are positioned at lower positions or upper positions. Then, the culture containers U20, U30are mounted on the swing plate33, whereby the culture containers U20, U30are accommodated in the culture container accommodating shelf32.

In the present embodiment, two swing plates33are attached to side edges which face four swing arms34supporting the swing plates33. Then, the swing arms34are turned to swing the swing plates33, thereby generating tilt to a horizontal plane in culture containers U20, U30, but a specific constitution of the swing plate33is not limited to this example. The culture containers U20, U30may only be tilted with a height difference so that the ports U21, U31are disposed at the lower positions or upper positions in accordance with the positions of the ports U21, U31disposed in the culture containers U20, U30.

It is to be noted that in the present embodiment, the swing plates33supported by the swing arms34are provided as the tilting means, but the culture containers U20, U30may be mounted on a plate which is constituted so as to turn and tilt in a predetermined periaxial direction and this plate may be defined as tilting means, as long as the culture containers U20, U30can be tilted as described above.

Alternatively, as shown inFIG. 6toFIG. 9, in the swing plate33, a stirring roller36to stir the content of the second culture container U30may be disposed as required. There is not any special restriction on a specific constitution of the stirring roller36, as long as the stirring roller pressed onto an upper surface of the second culture container U30reciprocatingly moves and can stir the content while bending and deforming the second culture container U30made of the soft packaging member.

It is to be noted that inFIG. 4, illustration of the stirring roller is omitted.

Furthermore, in removing the bubbles which enter into the first culture container U20or the second culture container U30from its port U21, U31, the removed bubbles can be transferred through the transfer tube U40to flow into the medium container U10, by releasing, for example, the closing with the closing member U44between the first culture container U20or the second culture container U30and the medium container U10to open the flow channel, and operating the pump U41. At this time, for the purpose of inhibiting the content of the first culture container U20or the second culture container U30from also flowing into the medium container U10together with the bubbles, the medium container accommodating shelf42in which the medium container U10is accommodated comprises weight detecting means43for detecting a weight change of the medium container U10in the present embodiment (seeFIG. 5andFIG. 10).

That is, when the content of the first culture container U20or the second culture container U30passes through the transfer tube U40together with the bubbles and is transferred to the medium container U10side, the weight increases due to the content. When the weight increase at this time is detected and the pump U41is automatically or manually stopped to end a transfer operation of the bubbles, the contents of the culture containers U20, U30can be inhibited from flowing into the medium container U10. Then, the pump U41is operated in reverse, whereby the content remaining in the transfer tube U40can be returned to the culture containers U20, U30and the content is not wasted.

It is to be noted that a period from when the weight increase is detected until the pump U41stops and a period for which the pump U41is operated in reverse can suitably be set in accordance with a volume of the transfer tube U40, a liquid delivery rate of the pump U41and the like.

Furthermore, in the present embodiment, the medium container accommodating shelf42comprises the weight detecting means43, but when the culture container accommodating shelf32comprises weight detecting means to detect a weight decrease when the content of the first culture container U20or the second culture container U30flows outside, thereby automatically or manually stopping the transfer operation of the bubbles, the contents of the culture containers U20, U30can be inhibited from flowing into the medium container U10.

Furthermore, in the present embodiment, the medium container U10is placed on a tray44, and the medium container U10is accommodated in the medium container accommodating shelf42via the tray44. That is, the medium container U10is not mounted directly on the medium container accommodating shelf42, but the medium container U10is placed on the tray44and is accommodated together with the tray44in the medium container accommodating shelf42.

Consequently, for example, when performing an operation of storing the cell culture unit U in the cell culture apparatus1, the first culture container U20and the second culture container U30can be placed on the tray44together with the medium container U10and then they can be carried. In consequence, the tray44is disposed on the medium container accommodating shelf42to accommodate the medium container U10, and then the first culture container U20and the second culture container U30can be taken out from the tray44and can be mounted on the culture container accommodating shelf32, whereby these series of operation can efficiently be performed.

Furthermore, in accommodating the medium container U10in the medium container accommodating shelf42via the tray44, it is preferable to tilt the tray44so that a side on which the port U11of the medium container U10is disposed becomes lower as shown inFIG. 10. Consequently, when the medium is supplied from the medium container U10to the culture containers U20, U30, the medium can be inhibited from remaining in the medium container U10.

The present invention has been described above with reference to the preferable embodiment, but needless to say, the present invention is not limited to the above-mentioned embodiment, and it is possible to implement various modifications in the gist of the present invention.

For example, the cell culture unit U to be stored in the cell culture apparatus1is not limited to the above-mentioned example. The unit may only comprise a culture container into which, together with a cell (a culture cell) that is a culture target, a medium prepared to culture the cell is supplied and the cell culture is performed, and a medium container to supply the medium to the culture container.

Alternatively, there is not any special restriction on the number of the medium containers to be connected, and the cell culture unit U can be constituted by connecting at least one medium container to the transfer tube. Furthermore, the cell culture unit U can be constituted by connecting, to the transfer tube, a collecting container to collect a content (a cell suspension) of the culture container in which the cell culture is ended.

Furthermore, the above-mentioned embodiment, there has been described the example where the inner portion of the casing of the cell culture apparatus1has the double tank structure divided into the incubating tank3and the cold insulation tank4by the insulation wall20, but there is not any special restriction on a specific constitution of each tank3or4, as long as the culture container accommodating shelf32and the medium container accommodating shelf42form a pair and are disposed separately in the incubating tank3and the cold insulation tank4. For example, although not especially shown in the drawing, the cell culture apparatus1may have a constitution in which a casing forming the incubating tank3and a casing forming the cold insulation tank4are independently disposed or these casings are coupled.

There are invoked herein all contents of the literature described in this description and the Japanese application description on the basis of which the priority of the present application under the Paris convention is claimed.

INDUSTRIAL APPLICABILITY

The present invention is suitably utilizable in a regenerative therapy, immunotherapy, antibody medicine production or the like to culture a large amount of cells in a closed system.

REFERENCE SIGNS LIST

1cell culture apparatus

30incubating tank side support frame

40cold insulation tank side support frame

43weight detecting means

U cell culture unit