JP2010075115A - Cell culture module - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a cell coupling module that does not perform an enzyme treatment such as a trypsin treatment, etc., easily releases cells cultured in a cell culture support and easily carries out a stepwise cell culture by using a plurality of cell culture supports since a movement operation by pipetting, etc. is not needed. <P>SOLUTION: The cell coupling module includes: a first sealing system member for sealably holding a first cell culture support arranged in a first recess; a first water pressure supply part for supplying a first water pressure to a second surface of the first cell culture support; a second sealing system member for sealably holding a second cell culture support arranged in a second recess larger than the first recess; a second water pressure supply part for supplying a second water pressure to a fourth surface of the second cell culture support; a cell conveying part for conveying cells released from the first recess from a first internal space to a second internal space; and a cell recovery part for recovering cells released from the second recess by the second water pressure from the second internal space. <P>COPYRIGHT: (C)2010,JPO&INPIT

Description

  The present invention relates to a cell culture module.

  In recent years, with the advancement of separation of stem cells from human bone marrow fluid, differentiation / induction to target tissue cells, 3D culture technology, development of scaffold materials, etc., cell cultures from stem cells to skin, bone, cartilage, blood vessels, Tissues such as heart valves and ligaments can be produced, and some clinical applications have already started.

  For such cell culture, a technique using a cell culture chip having a cell culture cell that is regularly arranged in an array shape or a honeycomb shape on the surface of a substrate and aggregates and holds seed cells is known. (For example, Patent Document 1).

Further, a cell culture device is known that includes a culture container that stores a culture solution containing a cell culture carrier and cells, and a magnetic field generator that generates a magnetic field for moving the cell culture carrier in the culture solution ( For example, Patent Document 2).
JP-A-2005-27598 JP 2004-313007 A

  In addition, the cell culture techniques as shown in Patent Documents 1 and 2 described above require a peeling operation from the cell culture chip or the cell culture carrier after cells are cultured. This peeling operation is usually performed using an enzyme treatment such as trypsin treatment.

  However, when such an enzyme treatment is carried out for a long time, the cultured cells deteriorate to some extent, and in the worst case, the cells themselves may die. In addition, enzyme treatment such as trypsin treatment has a problem that the operation is very complicated.

  In addition, when cultivating spherical cells, if the isolated cells are cultured suddenly in a large space, the cultured cells will not recognize the bottom of the space as a sphere, but as a flat surface. Is attached to the bottom, and spherical cells cannot be cultured. Therefore, in order to culture a spherical cell to a desired size, it is necessary to first culture the cell to a desired size in a small space, and then gradually culture the cells in a large space. That is, cells cultured on a cell culture carrier having a certain size of cell culture cell (hereinafter referred to as a recess) are transferred to another cell culture carrier having a larger recess and cultured in stages. It is necessary to continue.

  Conventionally, the means for moving such cultured cells to another cell culture carrier has been performed by pipetting after peeling by the above-described trypsin treatment, etc. It was cumbersome and inefficient.

  Therefore, the present invention has been made in view of the above problems, and cells cultured on a cell culture carrier can be easily detached from the cell culture carrier without performing enzyme treatment such as trypsin treatment, It is another object of the present invention to provide a cell culture module that can easily perform stepwise cell culture using a plurality of cell culture carriers because it is not necessary to perform a moving operation such as pipetting.

  The cell culture module according to the present invention includes a first closed system member that holds a first cell culture carrier composed of a porous body provided with a first recess for culturing cells in a first internal space so as to be capable of being sealed. In a state where the first inner space is filled with the culture solution, the first inner surface of the first cell culture carrier held in the first inner space is opposed to the first surface provided with the first recess. A first water pressure supply unit that supplies a first water pressure to the second surface; and a second recess that is larger than the first recess for culturing cells. A second sealing system member that holds the second cell culture carrier composed of the porous body in the second internal space in a sealable manner, and the second internal space is filled with a culture solution, On the third surface provided with the second recess of the second cell culture carrier held in the internal space of 2 A second water pressure supply unit that supplies a second water pressure to the fourth surface facing the second cell culture unit, and the first internal space and the second internal space are connected to each other And a cell transport unit that transports cells detached from the first recess by the first water pressure from the first internal space to the second internal space, and the cell transport unit at a position different from the cell transport unit. And a cell recovery unit that is connected to the second internal space and recovers the cells detached from the second recess by the second hydraulic pressure from the second internal space.

  By adopting such a configuration, the cells cultured on the cell culture carrier can be easily detached from the cell culture carrier without performing an enzyme treatment such as trypsin treatment. Therefore, stepped cell culture using a plurality of cell culture carriers can be easily performed.

  In addition, the cell culture module according to the present invention includes a first sealed system that holds a first cell culture carrier made of a porous body provided with a first recess for culturing cells in a first internal space so as to be hermetically sealed. A member, and the first recess and the first recess of the first cell culture carrier held in the first internal space in a state where the culture solution is filled in the first internal space. A first water pressure supply unit that supplies a first water pressure to the second surface opposite to the first surface, and a first cell culture unit including the first recess for culturing cells. A second sealing system member that holds the second cell culture carrier made of a porous body provided with a larger second concave portion in the second internal space in a sealable manner, and a culture solution in the second internal space. The second cell culture carrier of the second cell culture carrier held in the second internal space in a packed state. A second water pressure supply unit for supplying a second water pressure to a fourth surface opposite to a third surface provided with a second portion and the second recess, and the second cell culture unit, The first internal space and the second internal space are connected, and the cells detached from the first recess by the first hydraulic pressure with respect to the second surface of the first cell culture carrier are the first A cell transport unit transported from the internal space to the second internal space, the cell transport unit connected to the second internal space at a position different from the cell transport unit, and the fourth surface of the second cell culture carrier with respect to the fourth surface And a cell recovery section for recovering cells detached from the second recess by the second water pressure from the second internal space.

  With such a configuration, cells resistant to pressure such as chondrocytes can be cultured, and cells cultured on a cell culture carrier without performing enzyme treatment such as trypsin treatment can be used. In addition, since it is not necessary to perform a moving operation such as pipetting, stepwise cell culture using a plurality of cell culture carriers can be easily performed.

  It is preferable that a culture medium circulation system is further provided for filling one or both of the first internal space and the second internal space with a culture medium and circulating the filled culture medium. .

  By adopting such a configuration, in addition to the above-described effects, perfusion culture is also possible in the internal space provided with the culture medium circulation system.

  The connection part connected to the second internal space of the cell transfer part may be provided immediately above the second recess of the second cell culture carrier held in the second internal space. preferable.

  By setting it as such a structure, the cell conveyed from 1st internal space to 2nd internal space can be easily guide | induced to the 2nd recessed part of a 2nd cell culture support | carrier.

  More preferably, the second internal space is newly provided with cell rectification means for guiding the cells transferred from the cell transfer section into the second recesses of the second cell culture carrier. Preferably it is. Specifically, the cell rectifying means herein is a rectifying plate provided in the second internal space.

  By adopting such a configuration, the cells transported from the first internal space to the second internal space can be more reliably guided into the second recess of the second cell culture carrier.

  It is preferable that the size of the space constituting the second recess is equal and larger in all directions than the size of the space constituting the first recess.

  By setting it as such a structure, a spherical cell can be culture | cultivated reliably.

  In the present invention, cells cultured on a cell culture carrier can be easily detached from the cell culture carrier without performing an enzyme treatment such as trypsin treatment, and there is no need to perform a transfer operation such as pipetting. Therefore, a cell culture module capable of easily performing stepwise cell culture using a plurality of cell culture carriers is provided.

  Hereinafter, the present invention will be described in detail with reference to the drawings.

(First embodiment)
FIG. 1 is a conceptual diagram showing a cross-sectional configuration of the cell culture module according to the first embodiment. As shown in FIG. 1, the cell culture module 100 according to the present embodiment includes a first cell culture unit 101, a second cell culture unit 102, a cell transport unit 50, and a cell recovery unit 30. ing.

  The first cell culture unit 101 includes a first closed system member 10 and a first hydraulic pressure supply unit 20.

  For example, as shown in FIG. 1, the first sealing system member 10 includes a lower lid 10A, an upper lid 10B provided on the lower lid 10A, and a fixing member 15 that fixes the lower lid 10A and the upper lid 10B. It is configured.

Lower lid 10A, as shown in FIG. 1, to hold the first cell culture support S _1, and is configured in a concave shape with a recess 10Aa for receiving a culture medium.

  As shown in FIG. 1, the upper lid 10B is provided on the lower lid 10A, forms the lower lid 10A and the first inner space 12, and holds the first inner space 12 in a sealable manner. Convex part 10Ba which engages with concave part 10Aa of 10A so that sealing is possible is provided.

First holding cell culture carrier _{S 1,} for example, as shown in FIG. 1, the bottom portion 10Aa1 of recesses 10Aa of the lower cover 10A ○ - Place the ring 17a, ○ - first cell culture on the ring 17a the carrier _{S 1} is arranged, the first upper cell culture carrier _{S 1} ○ - place the ring 17b, ○ - ring 17a, the first cell culture carrier _{S 1} and ○ - ring 17b, the lower cover 10A and the upper cover and 10B when fixing together, the convex portion 10Ba of the upper cover 10B, ○ - by pressurizing the ring 17b, to retain the first cell culture support _{S 1} in the recess 10Aa of the lower cover 10A.

  In addition, when engaging the convex portion 10Ba of the upper lid 10B with the concave portion 10Aa of the lower lid 10A, the depression provided in the convex portion 10Ba of the upper lid 10B so that the first inner space 12 can be sealed. This is performed after attaching the O-ring 17c.

  The lower lid 10A and the upper lid 10B are made of, for example, acrylic resin.

  The fixing member 15 is made of, for example, SUS304.

  -The rings 17a, 17b, and 17c are made of, for example, fluorine rubber.

In the present embodiment, as described above, the first sealing member 10 is described as having a structure in which the lower lid 10A and the upper lid 10B are integrally fixed by the fixing member 15. being able to sealably retaining the first cell culture support S ₁ into the first internal space 12 is not limited to the above-described configuration.

First the first cell culture carrier S ₁ which is sealably retained in an internal space 12, the porous body in which the first recess S ₁₁ for culturing cells on the surface (first surface) Sa1 is provided It consists of

The first cell culture carrier S ₁ is configured zirconia, yttria, titania, alumina, silica, at least one kind of ceramics or glass of hydroxyapatite and β- tricalcium phosphate. These ceramics or glass are suitable because of high biostability.

  The first hydraulic pressure supply unit 20 is provided integrally or detachably on the bottom 10 </ b> Aa <b> 1 of the lower lid 10 </ b> A, and supplies the culture solution to the first internal space 12.

  As shown in FIG. 1, for example, the first hydraulic pressure supply unit 20 includes a culture solution storage unit 22 that stores a culture solution, one end attached to the bottom 10Aa1, and the other end attached to the storage unit 22. A culture medium supply pipe 24 that supplies the culture medium accommodated in the first internal space 12 to the first internal space 12 and a supply that is attached to the supply pipe 24 and controls the supply amount of the culture liquid supplied to the first internal space 12 An amount control unit 26 and a supply pressure control unit 28 that is attached to the storage unit 22 and controls the supply pressure when the culture solution is supplied.

  The culture solution storage unit 22 is composed of, for example, a resin storage container.

  The culture solution supply tube 24 is configured by, for example, a resin tube.

  The supply amount control unit 26 includes, for example, an electromagnetic valve that controls the supply amount of the culture solution. The opening / closing operation and the opening / closing amount of the electromagnetic valve are controlled by a control unit (not shown), and the supply amount of the culture solution is controlled by the opening / closing amount.

  The supply pressure control unit 28 includes, for example, a high-pressure gas supply device that supplies high-pressure gas from the culture solution storage unit 22 to the culture solution supply pipe 24.

Further, the first hydraulic supply unit 20, a first first first recess _{S 11} of the back side Sb1 (first cell culture carrier _{S 1} of the cell culture carrier _{S 1} held in the inner space 12 is The first water pressure is supplied to the second surface Sb1 by applying a hydraulic pressure in the direction of the surface (second surface) facing the provided surface (first surface) Sa1.

  The first water pressure is applied by supplying the supply amount controller 26 and the supply pressure so that the culture solution is supplied to the first internal space 12 in a state where the closed first internal space 12 is filled with the culture solution. This is done by controlling the control unit 28.

That is, the first internal space 12 is filled with the culture solution, the supply amount control unit 26 is set to the “open” state, and the supply pressure control unit 28 increases the supply pressure of the culture solution to increase the first By reducing the supply pressure, the water pressure is supplied to the second surface Sb1 of the _first cell culture carrier S1.

Wherein by applying a first pressure to the first of the second surface Sb1 cell culture carriers S _1, cells cultured in the first of the first recess S ₁₁ of the cell culture carrier S _1, a first cell It is peeled off from the culture carrier S ₁ and floats in the culture solution filled in the first internal space 12.

Incidentally, the first hydraulic pressure supplied to the first of the second surface Sb1 cell culture carriers S _1, in order to transfer to the surface of the first cell culture carrier S first recess S _{11 1,} first cell culture carrier S ₁ of is required to be composed of a porous body (porous body).

More preferably, the average pore diameter of the pores of the porous body constituting the first cell culture carrier S ₁ is is at 10nm or more 10μm or less, the pores of the first recess of the first cell culture carrier S ₁ from the surface of the S _11, it is preferably provided in communication to said second surface Sb1.

With such a configuration, transmitting the first pressure supplied to the first of the second surface Sb1 cell culture carriers S _1, to the surface of the first cell culture carrier S first recess S ₁₁ of ₁ it is possible to, the cells cultured in the first first recess S ₁₁ of the cell culture carrier S _1, can be easily peeled off from the first cell culture support S _1.

  The second cell culture unit 102 includes a second closed system member 110 and a second hydraulic pressure supply unit 120.

  For example, as shown in FIG. 1, the second sealing member 110 includes a lower lid 110A, an upper lid 110B provided on the lower lid 110A, and a fixing member 115 that fixes the lower lid 110A and the upper lid 110B. It is configured.

As shown in FIG. 1, the lower lid 110 </ b> A has a concave shape including a concave portion 110 </ b> Aa for holding the second cell culture carrier S <b> ₂ and accommodating the culture solution.

  As shown in FIG. 1, the upper lid 110B is provided on the lower lid 110A, forms the lower lid 110A and the second inner space 112, and holds the second inner space 112 in a sealable manner. Convex part 110Ba which engages with concave part 110Aa of 110A so that sealing is possible is provided.

Second holding cell culture carrier _{S 2,} for example, as shown in FIG. 1, the bottom 110Aa1 recess 110Aa of the lower cover 110A ○ - Place the ring 117a, ○ - second cell culture on the ring 117a the support _{S 2} is arranged on the second carrier for cell culture _{S 2} ○ - place the ring 117b, ○ - ring 117a, second cell culture support _{S 2} and ○ - ring 117b, the lower lid 110A and top lid and 110B when fixing together, the convex portion 110Ba of the upper lid 110B, ○ - by pressurizing the ring 117b, holding the second cell culture support _{S 2} in the recess 110Aa of the lower cover 110A.

  In addition, when engaging the convex portion 110Ba of the upper lid 110B with the concave portion 110Aa of the lower lid 110A, the depression provided in the convex portion 110Ba of the upper lid 110B so that the second inner space 112 is hermetically held. This is performed after attaching the ◯ -ring 117c to the.

  The lower lid 110A and the upper lid 110B are made of acrylic resin, for example.

  The fixing member 115 is made of, for example, SUS304.

  -The rings 117a, 117b, and 117c are made of, for example, fluorine-based rubber.

In the present embodiment, the second sealing system member 110 is described as having a structure in which the lower lid 110A and the upper lid 110B are integrally fixed by the fixing member 115, as described above. being able to sealably retaining the second cell culture support S ₂ to the second internal space 112 is not limited to the above-described configuration.

Second cell culture support S ₂ is sealably retained in the second internal space 112, porous body second recess S ₂₂ for culturing cells on the surface (third surface) Sa2 is provided It consists of The size of the space constituting the second concave portion S ₂₂ is configured larger than the size of the space constituting the first concave portion S _11.

The second cell culture support S ₂ is comprised of zirconia, yttria, titania, alumina, silica, at least one kind of ceramics or glass of hydroxyapatite and β- tricalcium phosphate. These ceramics or glass are suitable because of high biostability.

  The second hydraulic pressure supply unit 120 is provided integrally or detachably on the bottom 110Aa1 of the lower lid 110A, and supplies the culture solution to the second internal space 112.

  As shown in FIG. 1, for example, the second hydraulic pressure supply unit 120 includes a culture solution storage unit 122 that stores a culture solution, one end attached to the bottom portion 110Aa1, and the other end attached to the storage unit 122. A culture solution supply pipe 124 that supplies the culture solution accommodated in 122 to the second internal space 112, and a supply that is attached to the supply pipe 124 and controls the supply amount of the culture solution supplied to the second internal space 112 An amount control unit 126 and a supply pressure control unit 128 that is attached to the storage unit 122 and controls the supply pressure when the culture solution is supplied.

  The culture solution storage unit 122 is formed of, for example, a resin storage container.

  The culture solution supply pipe 124 is made of, for example, a resin tube.

  The supply amount control unit 126 includes, for example, an electromagnetic valve that controls the supply amount of the culture solution. The opening / closing operation and the opening / closing amount of the electromagnetic valve are controlled by a control unit (not shown), and the supply amount of the culture solution is controlled by the opening / closing amount.

  The supply pressure control unit 128 includes, for example, a high-pressure gas supply device that supplies high-pressure gas from the culture solution storage unit 122 to the culture solution supply pipe 124.

Further, the second hydraulic supply unit 120, a second second recess _{S 22} of the back surface of the cell culture carrier _{S 2} side Sb2 (second cell culture support _{S 2} is held in the second internal space 112 A second hydraulic pressure is supplied to the fourth surface Sb2 by applying a hydraulic pressure in the direction of the surface (fourth surface) facing the provided surface (third surface) Sa2.

  The method for applying the second water pressure includes the supply amount controller 126 and the supply pressure so that the culture solution is supplied to the second internal space 112 in a state where the sealed second internal space 112 is filled with the culture solution. This is done by controlling the control unit 128.

That is, in a state where the second internal space 112 is filled with the culture solution, the supply amount control unit 126 is set to the “open” state, and the supply pressure of the culture solution is increased by the supply pressure control unit 128. By reducing the supply pressure, the second water pressure is supplied to the fourth surface Sb2 of the _second cell culture carrier S2.

By providing the second water pressure on the fourth surface Sb2 of the second cell culture support S _2, cells cultured in the second of the second recess S ₂₂ cell culture carrier S ₂ is the second is peeled from the carrier for cell culture S _2, floating in the second have been culture in filling the inner space 112.

Incidentally, the second water pressure supplied to the second fourth surface Sb2 cell culture carrier S _2, in order to transfer to the surface of the second of the second recess S ₂₂ cell culture carrier S _2, a second the carrier for cell culture S ₂ it is necessary to be composed of a porous body (porous body).

More preferably, the average pore diameter of the pores of the porous body constituting the second cell culture support S ₂ is at 10nm or more 10μm or less, the pores said second of said second recess in the cell culture carrier S ₂ from the surface of the S _22, it is preferably provided in communication to said fourth surface Sb2.

With such a configuration, transmitting a second pressure which is supplied to the second fourth surface Sb2 cell culture carrier S _2, to the surface of the second of the second recess S ₂₂ cell culture carrier S ₂ it is possible to, the cells cultured in the second of the second recess S ₂₂ cell culture carrier S _2, can be easily peeled from the second cell culture support S _2.

Cells conveying unit 50 includes a first internal space 12 and a second internal space 112 is connected, the said the first hydraulic for the first of the second surface Sb1 cell culture carriers S ₁ first recess S _The cells detached from ₁₁ are conveyed from the first internal space 12 to the second internal space 112 together with the culture solution.

For example, as shown in FIG. 1, the cell transport unit 50 connects the upper portion 10Ba1 of the upper lid 10B of the first sealing member 10 and the upper portion 110Ba1 of the upper lid 110B of the second sealing member 110. a tube 52, attached to the carrier tube 52, controlling the conveyance amount of the culture solution containing the exfoliated cells from said first recess S ₁₁ from the first internal space 12 to the second internal space 112 And a carry amount control unit 54.

  The cell transfer tube 52 is made of, for example, a resin tube.

The transport amount control unit 54 is configured by, for example, an electromagnetic valve that controls the transport amount of the culture solution containing the detached cells. The solenoid valve control unit (not shown), the opening and closing operations and the opening and closing amount is controlled by opening and closing amounts, the conveyance amount of the culture solution containing the cells detached from the first recess S ₁₁ is controlled.

Cell recovery unit 30 (in FIG. 1, a second horizontal carrier for cell culture S ₂₎ cell conveying pipe 52 different from the location of cells conveying portion 50 integral with the second closed system member 110 or removably connected to the second internal space 112, cultured in the second of the second recess S ₂₂ cell culture carrier S _2, a second water pressure supplied to the second fourth surface Sb2 cell culture carrier S ₂ , to recover the cells detached from the second cell culture carrier S ₂ from the second internal space 112 per culture.

  For example, as shown in FIG. 1, the cell recovery unit 30 includes a cell storage unit 32 that stores cultured cells together with a culture solution, one end attached to the second sealing member 110, and the other end attached to the storage unit 32. A cell discharge pipe 34 for discharging the cells cultured in the second internal space 112 together with the culture solution from the internal space 112, and discharge of the culture solution attached to the discharge pipe 34 and discharged from the internal space 112 It is comprised with the discharge amount control part 36 which controls quantity.

  The cell accommodating part 32 is comprised by the resin-type accommodation container, for example.

  The cell discharge tube 34 is composed of, for example, a resin tube.

  The discharge amount control unit 36 is configured by, for example, an electromagnetic valve that controls the discharge amount. The opening / closing operation and opening / closing amount of the solenoid valve are controlled by a control unit (not shown), and the discharge amount is controlled by the opening / closing amount.

  Next, an example of a cell culture method using the cell culture module 100 shown in FIG. 1 will be described with reference to FIGS.

  2 to 4 are conceptual diagrams for explaining an example of a cell culture method using the cell culture module according to the first embodiment.

First, feeder cells (Feeder cells) are placed on the second surface Sb1 of the _first cell culture carrier S1.

Then, prepare a concave lower lid 10A, the first cell culture carrier _{S 1} and O- ring 17b of the bottom 10Aa1 the O- ring 17a of the lower cover 10A, feeder cells (Feeder cells) is installed They are stacked in order and placed in the recess 10Aa of the lower lid 10A. Then, placing the cells to be cultured in the first of the first recess _{S 11} of the cell culture carrier _{S 1} which is placed in the recess 10Aa of the lower cover 10A. Incidentally, when the cell to be cultured are placed in the first of the first recess S ₁₁ of the cell culture carrier S _1, the installation cells are attached to the first cell culture carrier S ₁ surface, it is retained .

  Thereafter, the O-ring 17c is inserted into the recess of the convex portion 10Ba of the upper lid 10B, and the concave portion 10Aa of the lower lid 10A and the convex portion 10Ba of the upper lid 10B are fitted to each other, and the lower lid 10A is fixed by the fixing member 15. And the upper lid 10 </ b> B are integrally fixed to constitute the first sealed system member 10 according to the present embodiment in which the first internal space 12 is formed.

Then, the 1st cell culture part 101 which concerns on this embodiment is comprised by attaching the culture solution supply pipe | tube 24 of the 1st hydraulic pressure supply part 20 to bottom part 10Aa1 of 10 A of lower lids.

Next, the second closed system member 110 is configured in the same manner as the first closed system member 10. That is, first, feeder cells (Feeder cells) are placed on the fourth surface Sb2 of the _second cell culture carrier S2.

Then, prepare a concave lower lid 110A, the bottom 110Aa1 the O- rings 117a, feeder cells (Feeder cells) second cell culture support _{S 2} and O- ring 117b disposed in the lower lid 110A They are stacked in order and placed in the recess 110Aa of the lower lid 110A.

  Thereafter, the O-ring 117c is inserted into the recess of the convex portion 110Ba of the upper lid 110B, and the concave portion 110Aa of the lower lid 110A and the convex portion 110Ba of the upper lid 110B are fitted to each other, and the lower lid 110A is fixed by the fixing member 115. And the upper lid 110 </ b> B are integrally fixed to constitute the second sealed member 110 according to the present embodiment in which the second internal space 112 is formed.

  Thereafter, the second cell culture unit 102 according to the present embodiment is configured by attaching the culture solution supply pipe 124 of the second hydraulic pressure supply unit 120 to the bottom 110Aa1 of the lower lid 110A.

  Finally, one end of the cell transfer tube 52 of the cell transfer unit 50 is placed on the upper portion 10Ba1 of the upper lid 10B of the first sealing member 10 and the other end is placed on the upper portion 110Ba1 of the upper lid 110B of the second sealing member 110, respectively. The cell culture module 100 which concerns on this embodiment is comprised by connecting the cell discharge pipe 34 of the cell collection | recovery part 30 with the 2nd sealing system member 110 in the position different from the said cell conveyance pipe | tube 52 while connecting.

  Next, the discharge amount control unit 36 is set to the “open” state (in this embodiment, the solenoid valve of the discharge amount control unit 36 is set to the “open” state), and the transport amount control unit 54 is set to the “open” state (this embodiment). Then, the electromagnetic valve of the conveyance amount control unit 54 is set to the “open” state, and the supply amount control units 26 and 126 are set to the “open” state (in this embodiment, the electromagnetic valves of the supply amount control units 26 and 126 are set to “ `` Open '' state), by applying the supply pressure from the supply pressure control unit 28, 128, until the culture solution is completely filled in the first internal space 12, the second internal space 112, and the cell transport tube 52, The culture solution is supplied from the culture solution storage units 22 and 122.

  When the culture solution is completely filled in the first internal space 12, the second internal space 112, and the cell transport pipe 52, the supply pressure from the supply pressure control units 28, 128 is stopped, and the supply amount control unit 26 126 are set to the “closed” state (in this embodiment, the solenoid valves of the supply amount control units 26 and 126 are set to the “closed” state), and at the same time, the discharge amount control unit 36 is set to the “closed” state (the discharge amount is set in this embodiment). The electromagnetic valve of the amount control unit 36 is set to the “closed” state, and the conveyance amount control unit 54 is set to the “closed” state (in this embodiment, the electromagnetic valve of the conveyance amount control unit 54 is set to the “closed” state).

Thus, the environment is ready to cells retained in the first first recess S ₁₁ of the cell culture carrier S ₁ is cultured in the first internal space 12 (FIG. 2).

After culturing the cells in the first internal space 12, the discharge amount control unit 36 is set to the “open” state to start the discharge of the culture solution in the second internal space 112, and the transport amount control unit 54 and the supply amount control unit 26 is set in the “open” state, and supply pressure is supplied from the supply pressure control unit 28 to start supplying a new culture solution to the first internal space 12. By this operation, the hydraulic pressure P ₁ is supplied from the first hydraulic pressure supply unit 20 in the direction of the second surface Sb1 of the _first cell culture carrier S1. The fluid pressure P _1, since the first of the second surface Sb1 cell culture carriers S ₁ pressure is supplied, cells cultured in the first of the first recess S ₁₁ of the cell culture carrier S ₁ is The first cell culture carrier S ₁ is peeled off from the surface of the first recess S ₁₁ , floats in the culture medium in the first internal space 12, and flows through the cell transport tube 52 together with the culture medium. , is conveyed to the second internal space 112, then into the second internal space 112 second second recess S ₂₂ cell culture carrier S ₂ held in the cell enters the second It is in the the recess S ₂₂ attached to the second surface of the cell culture carrier S _2, is maintained.

  Thereafter, after the supply pressure is stopped from the supply pressure control unit 28, the supply amount control unit 26, the discharge amount control unit 36, and the transport amount control unit 54 are set in the “closed” state.

Thus, the environment is ready to cells retained in the second of the second recess S ₂₂ cell culture carrier S ₂ are cultured in the second internal space 112 (FIG. 3).

After culturing cells in the second internal space 112, the discharge amount control unit 36 is set in the “open” state to start discharging the culture solution in the second internal space 112, and the supply amount control unit 126 is set in the “open” state. Furthermore, the supply pressure is supplied from the supply pressure control unit 128 to start supplying a new culture solution to the second internal space 112. This operation, in the second fourth surface Sb2 direction of the cell culture carrier _{S 2,} the hydraulic pressure _{P 2} is supplied from the second hydraulic supply unit 120. The fluid pressure P _2, since the water pressure in the second fourth surface Sb2 cell culture carrier S ₂ is supplied, cells cultured in the second of the second recess S ₂₂ cell culture carrier S ₂ is is peeled from the surface of the second of the second recess S ₂₂ cell culture carrier S _2, suspended in culture medium within the second internal space 112, it is, together with the culture solution, the cells discharge pipe of cell recovery section 30 Through 34, it is collected in the cell accommodating part 32 (FIG. 4).

As described above, since the cell culture module according to the present embodiment has the above-described configuration, cells cultured on the cell culture carrier can be easily removed from the cell culture carrier without performing enzyme treatment such as trypsin treatment. Can be peeled off. Further, since there is no need to perform a moving operation such as pipetting, stepwise cell culture using a plurality of cell culture carriers can be easily performed.

In addition, although the cell conveyance pipe | tube 52 demonstrated by this embodiment demonstrated the structure which connected one end to upper part 10Ba1 of the upper cover 10B of the 1st sealing system member 10 in FIG. 1, this invention is limited to this. For example, as shown in FIG. 5, one end of the cell transport tube 52 may be connected in the horizontal direction of the _first cell culture carrier S1 held in the recess 10Aa of the lower lid 10A. Good.

As shown in FIG. 1, the connecting part connected to the second internal space 112 of the cell transport unit 50 is the second recess S of the second cell culture carrier S ₂ held in the second internal space 112. It is preferable to be provided immediately above ₂₂ . With such a configuration, it is guided to the first internal space 12 from the second cells that have been transported in the interior space 112 facilitates the second second cell culture carrier S ₂ of the concave S ₂₂ it can.

More preferably, the second inner space 112, the new cell rectifying means for guiding the cells conveyed from the cell transport unit 50 in the second of the second recess S ₂₂ cell culture carrier S ₂ Is preferably provided.

With such a configuration, more reliably, the first internal space 12 from the second internal space 112 is conveyed to the cells easily in the second of the second recess S ₂₂ carrier for cell culture S ₂ Can lead to.

  Specifically, as shown in FIG. 6, such a cell rectifier means a rectifier attached to the inside of the convex portion 110Ba of the upper lid 110B of the second sealing member 110, that is, the second internal space 112. It is preferable that the plate 150 be configured in a ring shape (in FIG. 6, the inner diameter of the ring is reduced downward). 17A and 17B are a cross-sectional view and a top view of the rectifying plate 150. FIG. As shown in FIGS. 17A and 17B, the rectifying plate 150 has a ring shape, and the ring-shaped central portion 151a has a configuration in which the diameter decreases downward. Note that the rectifying plate 150 here is made of, for example, an acrylic resin.

Size of the space constituting the second concave portion S ₂₂ is preferably greater at the same magnification in all directions than the size of the space constituting the first concave portion S _11.

7, (a) a top view of the first of the first recess _{S 11} near the cell culture carriers _{S 1} are used for the cell culture module according to the first embodiment, A-A of (b) (a) a cross-sectional view taken along the line 8, (a) a top view of the second of the second recess S ₂₂ near the cell culture carrier S ₂ are used for the cell culture module of the first embodiment, (b) ( It is sectional drawing in the BB line of a).

Here, “the same size and larger in all directions” means that both the width X ₂ and the depth Y ₂ of the opening on the upper surface of the second recess S ₂₂ shown in FIG. greater at the same magnification the same than the width _{X 1} and the depth _{Y 1} of the opening of the upper surface of the concave portion _{S 11, i.e., X 2 = aX 1, Y} 2 = aY 1 (a is greater than 1 natural number) is composed of It means being.

  By setting it as such a structure, a spherical cell can be culture | cultivated reliably.

(Second Embodiment)
FIG. 9 is a conceptual diagram showing a cross-sectional configuration of the cell culture module according to the second embodiment. In the cell culture module 200 according to the present embodiment, the first cell culture unit 101 shown in FIG. 5 described in the first embodiment is provided with a third hydraulic pressure supply unit 60. The difference is that the second cell culture unit 102 is replaced with a second cell culture unit 102a newly provided with a fourth hydraulic pressure supply unit 160. Since others are the same as those of the first embodiment, description thereof is omitted.

  The third hydraulic pressure supply unit 60 is provided integrally or detachably on the upper portion 10Ba1 of the upper lid 10B of the first sealing system member 10, and supplies the culture solution to the first internal space 12.

  For example, as shown in FIG. 9, the third hydraulic pressure supply unit 60 includes a culture solution storage unit 62 that stores a culture solution, one end attached to the upper portion 10Ba1 of the upper lid 10B, and the other end attached to the storage unit 62. A culture solution supply pipe 64 that supplies the culture solution stored in the storage portion 62 to the first internal space 12, and a supply that is attached to the supply tube 64 and controls the supply amount of the culture solution supplied to the internal space 12 An amount control unit 66 and a supply pressure control unit 68 that is attached to the storage unit 62 and controls the supply pressure when the culture solution is supplied.

  The culture solution storage unit 62 is formed of, for example, a resin storage container.

  The culture solution supply pipe 64 is made of, for example, a resin tube.

  The supply amount control unit 66 includes, for example, an electromagnetic valve that controls the supply amount of the culture solution. The opening / closing operation and the opening / closing amount of the electromagnetic valve are controlled by a control unit (not shown), and the supply amount of the culture solution is controlled by the opening / closing amount.

The supply pressure control unit 68 includes, for example, a high-pressure gas supply device that supplies high-pressure gas from the culture solution storage unit 62 to the culture solution supply pipe 64.

The third hydraulic supply unit 60, by giving the fluid pressure in the first of the first recess S ₁₁ direction of the cell culture carrier S ₁ held in the first internal space 12, the first recess S ₁₁ is supplied with water pressure.

  In the method of applying the water pressure, the supply amount control unit 66 and the supply pressure control unit 68 are controlled so that the culture solution is supplied to the internal space 12 in a state where the sealed first internal space 12 is filled with the culture solution. To do.

That is, in a state where the first inner space 12 is filled with the culture solution, the supply amount control unit 66 is set to the “open” state, and the supply pressure of the culture solution is increased by the supply pressure control unit 68 to increase the water pressure. small water pressure by reducing the supply pressure is supplied to the first of the first recess S ₁₁ of the cell culture carrier S _1. While providing hydraulic pressure by the third pressure supply unit 60 to the first of the first recess S ₁₁ direction of the cell culture carrier S _1, when the cells are cultured in the first recess S _11, the pressure of such chondrocytes Strong cells can be cultured.

  The fourth hydraulic pressure supply unit 160 is provided integrally or detachably on the upper portion 110Ba1 of the upper lid 110B of the second sealing system member 110, and supplies the culture solution to the second internal space 112.

  For example, as shown in FIG. 9, the fourth hydraulic pressure supply unit 160 includes a culture solution storage unit 162 that stores a culture solution, and one end of a cell between the conveyance amount control unit 54 and the second sealed system member 110. The other end of the transfer tube 52 is attached to the storage unit 162, and the culture solution supply pipe 164 that supplies the culture solution stored in the storage unit 162 to the second internal space 112 is attached to the supply tube 164. A supply amount control unit 166 that controls the supply amount of the culture solution supplied to the internal space 112 and a supply pressure control unit 168 that is attached to the storage unit 162 and controls the supply pressure when supplying the culture solution.

  The culture solution storage unit 162 is formed of, for example, a resin storage container.

  The culture solution supply pipe 164 is made of, for example, a resin tube.

  The supply amount control unit 166 includes, for example, an electromagnetic valve that controls the supply amount of the culture solution. The opening / closing operation and the opening / closing amount of the electromagnetic valve are controlled by a control unit (not shown), and the supply amount of the culture solution is controlled by the opening / closing amount.

  The supply pressure control unit 168 includes, for example, a high pressure gas supply device that supplies high pressure gas from the culture solution storage unit 162 to the culture solution supply pipe 164.

The fourth pressure supply unit 160 to provide a fluid pressure to the second of the second recess S ₂₂ direction of the cell culture carrier S _2, which is held in the second internal space 112, the second concave portion S Water pressure is supplied to ₂₂ .

  In the method of applying the water pressure, the supply amount control unit 166 and the supply pressure control unit 168 are controlled so that the culture solution is supplied to the internal space 112 in a state where the sealed second internal space 112 is filled with the culture solution. To do.

That is, in a state where the second internal space 112 is filled with the culture solution, the supply amount control unit 166 is set to the “open” state, and the supply pressure of the culture solution is increased by the supply pressure control unit 168, thereby increasing the water pressure. small water pressure by reducing the supply pressure is supplied to the second of the second recess S ₂₂ of the cell culture carrier S _2. While applying the hydraulic pressure by the fourth pressure supply unit 160 to the second of the second recess S ₂₂ direction of the cell culture carrier S _2, when the cells are cultured in the second recess S _22, the second internal space 112 In the inside, cells resistant to pressure such as chondrocytes can be cultured.

  Next, an example of a cell culture method using the cell culture module 200 shown in FIG. 9 will be described with reference to FIGS. In addition, description of the part which overlaps with the part demonstrated in 1st Embodiment is abbreviate | omitted.

  10 to 13 are conceptual diagrams for explaining an example of a cell culture method using the cell culture module according to the second embodiment.

  First, the first closed system member 10 and the second closed system member 110 according to the present embodiment are configured.

  Thereafter, the culture solution supply pipe 24 of the first hydraulic pressure supply unit 20 is attached to the bottom 10Aa1 of the lower lid 10A of the first sealing system member 10, and a third 10Ba1 of the upper lid 10B of the first sealing system member 10 is attached to the third lid 10A1. By attaching the culture solution supply pipe 64 of the water pressure supply unit 60, the first cell culture unit 101a is configured.

Further, the culture solution supply pipe 124 of the second hydraulic pressure supply unit 120 is attached to the bottom part 110Aa1 of the lower lid 110A of the second sealing member 110, and one end of the cell transfer pipe 52 of the cell transfer part 50 is connected to the first part. a first horizontal carrier for cell culture _{S 1} of the closed system member 10, the other end is respectively connected to the top 110Ba1 the lid 110B of the second closed system members 110, furthermore, the cell discharge pipe of cell recovery section 30 34 the coupling to the second closed system member 110 in cell transport tube 52 a different position (in FIG. 9, a second horizontal carrier for cell culture S _2).

Finally, by connecting the culture medium supply pipe 164 of the transport amount control unit 54 and the fourth pressure supply unit 160 to the cell conveying pipe 52 through the connecting portion R ₁ between the second closed system member 110 The second cell culture unit 102a including the second closed system member 110, the second hydraulic pressure supply unit 120, and the fourth hydraulic pressure supply unit 160 is configured, and the first cell culture unit 101a and the second cell The cell culture module 200 according to the present embodiment including the culture unit 102a, the cell transport unit 50, and the cell recovery unit 30 is configured.

  Next, the discharge amount control unit 36 is set to the “open” state, the transport amount control unit 54 is set to the “open” state, and the supply amount control units 26 and 126 are set to the “open” state, respectively. By supplying a supply pressure from 128, the culture solution is supplied from the culture solution storage units 22, 122 until the culture solution is completely filled in the first internal space 12, the second internal space 112, and the cell transfer tube 52. To do.

  When the culture solution is completely filled in the first internal space 12, the second internal space 112, and the cell transport pipe 52, the supply pressure from the supply pressure control units 28, 128 is stopped, and the supply amount control unit 26 126 are set to the “closed” state, and at the same time, the discharge amount control unit 36 is set to the “closed” state, and the transport amount control unit 54 is set to the “closed” state.

Next, the supply amount control unit 66 is set to the “open” state, and the supply pressure is supplied from the supply pressure control unit 68, whereby the first cell culture carrier S ₁ held in the first internal space 12 is supplied with the first pressure. by providing a hydraulic _{P 3} into the recess _{S 11} direction, and supplies the water pressure to the first recess _{S 11.}

Thus, the environment is ready to cells retained in the first first recess S ₁₁ of the cell culture carrier S ₁ is cultured in the first internal space 12 (FIG. 10).

After cell culture in the first internal space 12, the supply pressure from the supply pressure control unit 68 is stopped, the supply amount control unit 66 is set in the “closed” state, and the discharge amount control unit 36 is set in the “open” state. The discharge of the culture solution in the internal space 112 is started, the conveyance amount control unit 54 and the supply amount control unit 26 are set to the “open” state, and supply pressure is supplied from the supply pressure control unit 28 to supply the first internal space. 12, the supply of the culture solution is newly started. By this operation, the hydraulic pressure P ₁ is supplied from the first hydraulic pressure supply unit 20 in the direction of the second surface Sb1 of the _first cell culture carrier S1. The fluid pressure P _1, since the first of the second surface Sb1 cell culture carriers S ₁ pressure is supplied, cells cultured in the first of the first recess S ₁₁ of the cell culture carrier S ₁ is The first cell culture carrier S ₁ is peeled off from the surface of the first recess S ₁₁ , floats in the culture medium in the first internal space 12, and flows through the cell transport tube 52 together with the culture medium. The cells enter the second recess S ₂₂ of the second cell culture carrier S ₂ that is transported into the second inner space 112 and held in the second inner space 112, and the second recess S within ₂₂ attached to the second surface of the cell culture carrier _{S 2,} it is held (Fig. 11).

  Thereafter, the supply pressure is stopped from the supply pressure control unit 28, and the supply amount control unit 26, the discharge amount control unit 36, and the transport amount control unit 54 are set to the “closed” state.

Then, the supply amount control unit 166 to "open" state, to provide a supply pressure from the supply pressure control section 168, a second cell culture carrier S ₂ second held in the second internal space 112 by providing a hydraulic _{P 4} into the recess _{S 22} direction, and supplies the water pressure to the second concave portion _{S 22.}

Thus, the environment is ready to cells retained in the second of the second recess S ₂₂ cell culture carrier S ₂ are cultured in the second internal space 112 (Fig. 12).

After cell culture in the second internal space 112, the supply pressure from the supply pressure control unit 168 is stopped, the supply amount control unit 166 is set in the “closed” state, and the discharge amount control unit 36 is set in the “open” state. The discharge of the culture solution in the internal space 112 is started, the supply amount control unit 126 is set to the “open” state, and the supply pressure is supplied from the supply pressure control unit 128 to newly add the culture solution to the second internal space 112. Start supplying. This operation, in the second fourth surface Sb2 direction of the cell culture carrier _{S 2,} the hydraulic pressure _{P 2} is supplied from the second hydraulic supply unit 120. The fluid pressure P _2, since the water pressure in the second fourth surface Sb2 cell culture carrier S ₂ is supplied, cells cultured in the second of the second recess S ₂₂ cell culture carrier S ₂ is is peeled from the surface of the second of the second recess S ₂₂ cell culture carrier S _2, suspended in culture medium within the second internal space 112, it is, together with the culture solution, the cells discharge pipe of cell recovery section 30 Through 34, it is collected in the cell accommodating part 32 (FIG. 13).

As described above, since the cell culture module according to the present embodiment has the above-described configuration, cells that are resistant to pressure such as chondrocytes can be cultured, and enzyme treatment such as trypsin treatment can be performed. In addition, the cells cultured on the cell culture carrier can be easily detached from the cell culture carrier. Further, since there is no need to perform a moving operation such as pipetting, stepwise cell culture using a plurality of cell culture carriers can be easily performed.

Note that the culture solution supply pipe 164 of the fourth hydraulic pressure supply unit 160 described in the present embodiment is connected to the cell transfer pipe 52 between the transfer amount control unit 54 and the second sealing system member 110 by connecting the connecting portion R ₁ . However, the present invention is not limited to this. For example, as shown in FIG. 14, the cell transfer tube 52 and the culture solution supply tube 164 are respectively attached to the upper portion 110Ba1 of the upper lid 110B. You may connect independently ((alpha) in FIG. 14). Such a configuration is preferable because the supply of the water pressure P ₄ from the fourth water pressure supply unit 160 becomes more efficient.

(Third embodiment)
FIG. 15 is a conceptual diagram showing a cross-sectional configuration of the cell culture module according to the third embodiment. In the cell culture module 300 according to the present embodiment, the first cell culture unit 101 shown in FIG. 5 described in the first embodiment newly includes a first culture medium circulation system 500. The difference is that the second cell culture unit 102 is replaced with the second cell culture unit 102b newly provided with the second culture medium circulation system 600. Since others are the same as that of 1st Embodiment, description is abbreviate | omitted.

  The first culture fluid circulation system 500 supplies the culture fluid to the first internal space 12 and circulates the culture fluid supplied to the first internal space 12. The second culture fluid circulation system 600 supplies the culture fluid to the second internal space 112 and circulates the culture fluid supplied to the second internal space 112.

For example, as shown in FIG. 15, the first culture fluid circulation system 500 holds a culture fluid in advance and temporarily stores the circulated culture fluid, and the accommodation portion 512. The supply device 514 that supplies the culture solution stored in the first internal space 12 from the circulation direction Dn, and the pipes 516a and 516b that connect the first sealing member 10, the storage portion 512, and the supply device 514. 516c, and circulation amount control units 518a and 518b that are attached to the pipes 516a and 516c and control ON / OFF of the circulation of the first culture solution circulation system 500 and the circulation amount thereof. Piping 516a is on the bottom 10Aa1 of the lower cover 10A of the first closed system member 10, the pipe 516c, a first carrier for cell culture _{S 1} first upper 10Ba1 of the top cover 10B of the first closed system member 10 They are respectively connected to right above the concave S _11.

  The culture solution storage unit 512 is formed of, for example, a resin storage container.

  The supply device 514 is constituted by a pump, for example.

  The pipes 516a, 516b, and 516c are made of, for example, a resin tube.

  The circulation amount control units 518a and 518b are configured by, for example, electromagnetic valves as described in the first embodiment.

As shown in FIG. 15, the culture fluid supply pipe 24 of the first hydraulic pressure supply unit 20 is connected to a pipe 516a between the circulation amount control unit 518a and the bottom 10Aa1 of the lower lid 10A of the first closed system member 10. It is connected via a section R _2.

For example, as shown in FIG. 15, the second culture fluid circulation system 600 holds the culture fluid in advance, and temporarily stores the circulated culture fluid, and the accommodation portion 612. The supply device 614 for supplying the culture solution stored in the second internal space 112 from the circulation direction Dn, and the piping 616a, 616b for connecting the second sealing system member 110, the storage portion 612, and the supply device 614 to each other. , 616c, and circulation amount control units 618a and 618b that are attached to the pipes 616a and 616c and control the circulation amount of the second culture medium circulation system 600. Piping 616a is on the bottom 110Aa1 of the second lower cover 110A of the closed system member 110, pipe 616c, the second of the second cell culture carrier _{S 2} of the upper 110Ba1 the lid 110B of the second closed system member 110 They are respectively connected to right above the concave S _22.

  The culture solution storage unit 612 is formed of, for example, a resin storage container.

  The supply device 614 is configured by, for example, a pump.

  The pipes 616a, 616b, 616c are made of, for example, a resin tube.

  The circulation amount control units 618a and 618b are configured by, for example, electromagnetic valves as described in the first embodiment.

As shown in FIG. 15, the culture fluid supply pipe 124 of the second hydraulic pressure supply unit 120 is connected to a pipe 616a between the circulation amount control unit 618a and the bottom 110Aa1 of the lower lid 110A of the second sealing system member 110. It is connected via a section R _3.

In the cell culture module 300 shown in the present embodiment, the supply of the hydraulic pressure P ₁ by the first hydraulic pressure supply unit 20 causes the circulation amount control units 518a and 518b of the first culture fluid circulation system 500 to be in a “closed” state. Others are performed by the same method as described in the first embodiment.

Also in the supply of hydraulic pressure _{P 2} of the second hydraulic supply unit 120, the circulation quantity control unit of the second culture solution circulation system 600 618a, the 618b, the "closed" state, the other is the first embodiment The method is the same as the method described in the embodiment.

Further, the supply of the hydraulic pressure _{P 3} as described above, the circulation quantity control unit of the first culture solution circulation system 500 518a, the 518b, as the state "open", the culture solution by supplying device 514 from the medium accommodating portion 512 first 1 is supplied into the internal space 12 and circulated from the circulation direction Dn.

In addition, the supply of the fluid pressure P ₄ described above causes the circulation amount control units 618 a and 618 b of the second culture solution circulation system 600 to be in the “open” state, and the culture solution is supplied from the culture solution storage unit 612 by the supply device 614. 2 is supplied into the internal space 112 and circulated from the circulation direction Dn.

As described above, since the cell culture module according to the present embodiment has the above-described configuration, cells that are resistant to pressure such as chondrocytes can be cultured, and enzyme treatment such as trypsin treatment can be performed. In addition, the cells cultured on the cell culture carrier can be easily detached from the cell culture carrier. Further, since there is no need to perform a moving operation such as pipetting, stepwise cell culture using a plurality of cell culture carriers can be easily performed.

Note that the culture solution supply pipe 24 of the first hydraulic pressure supply unit 20 described in the present embodiment is connected to a pipe 516a between the circulation amount control unit 518a and the bottom 10Aa1 of the lower lid 10A of the first closed system member 10. described configurations coupled through a coupling portion R _2, but the present invention is not limited thereto, for example, as shown in FIG. 16, the piping 516a and the culture solution supply pipe 24 of the lower cover 10A You may connect with bottom part 10Aa1 each independently ((beta) in FIG. 16). In addition, the culture solution supply pipe 124 of the second hydraulic pressure supply unit 120 described in the present embodiment is connected to a pipe 616a between the circulation amount control unit 618a and the bottom 110Aa1 of the lower lid 110A of the second sealing system member 110. described configurations coupled through a coupling portion R _3, but the present invention is not limited thereto, for example, as shown in FIG. 16, the piping 616a and the culture solution supply pipe 124 of the lower cover 110A You may connect with bottom part 110Aa1 each independently ((gamma) in FIG. 16).

Such a configuration is preferable because the supply of the water pressure P ₁ from the first water pressure supply unit 20 and the supply of the water pressure P ₂ from the second water pressure supply unit 120 become more efficient.

(Other embodiments)
As mentioned above, although embodiment of this invention was described in detail, this invention can perform various design change and improvement in the range which does not deviate from the summary.

For example, as described above, the cell culture modules 100 and 200 described in the first and second embodiments include the first culture medium circulation system 500 and the second culture medium circulation system 600 described in the third embodiment. May be further provided. In this case, the first culture fluid circulation system 500 and the second culture fluid circulation system 600 are placed in the first recess S ₁₁ and the second recess S ₂₂ as described in the third embodiment. ) May be provided, or may be arranged only for circulating the culture solution in the first internal space 12 and the second internal space 112.

  In addition, when arrange | positioning only for circulating the culture solution in the 1st internal space 12 and the 2nd internal space 112, the 1st culture solution circulation system 500 or the 2nd culture solution circulation system 600 is used. Only one of them may be arranged, or both the first culture solution circulation system 500 and the second culture solution circulation system 600 may be arranged.

  By adopting such a configuration, in addition to the above-described effects, perfusion culture is also possible in the internal space provided with the culture medium circulation system.

It is a conceptual diagram which shows the cross-sectional structure of the cell culture module which concerns on 1st Embodiment. It is a conceptual diagram for demonstrating an example of the cell culture method using the cell culture module which concerns on 1st Embodiment. It is a conceptual diagram for demonstrating an example of the cell culture method using the cell culture module which concerns on 1st Embodiment. It is a conceptual diagram for demonstrating an example of the cell culture method using the cell culture module which concerns on 1st Embodiment. It is a conceptual diagram which shows the cross-sectional structure of the other aspect of the cell culture module which concerns on 1st Embodiment. It is a conceptual diagram which shows the cross-sectional structure of the other aspect of the cell culture module which concerns on 1st Embodiment. (A) top view of a first of the first recess S ₁₁ near the cell culture carriers S ₁ are used for the cell culture module according to the first embodiment, cross-sectional view along line A-A of (b) (a) It is. (A) a top view of the second of the second recess S ₂₂ near the cell culture carrier S ₂ are used for the cell culture module of the first embodiment, in cross-section along the line B-B of (b) (a) is there. It is a conceptual diagram which shows the cross-sectional structure of the cell culture module which concerns on 2nd Embodiment. It is a conceptual diagram for demonstrating an example of the cell culture method using the cell culture module which concerns on 2nd Embodiment. It is a conceptual diagram for demonstrating an example of the cell culture method using the cell culture module which concerns on 2nd Embodiment. It is a conceptual diagram for demonstrating an example of the cell culture method using the cell culture module which concerns on 2nd Embodiment. It is a conceptual diagram for demonstrating an example of the cell culture method using the cell culture module which concerns on 2nd Embodiment. It is a conceptual diagram which shows the cross-sectional structure of the other aspect of the cell culture module which concerns on 2nd Embodiment. It is a conceptual diagram which shows the cross-sectional structure of the cell culture module which concerns on 3rd Embodiment. It is a conceptual diagram which shows the cross-sectional structure of the other aspect of the cell culture module which concerns on 3rd Embodiment. It is (a) sectional drawing of the baffle plate 150 used for the cell culture module which concerns on 1st Embodiment, (b) Top view.

Explanation of symbols

DESCRIPTION OF SYMBOLS 10 1st sealing system member 10A Lower lid 10B Upper lid 12 1st internal space 15 Fixing member 20 1st hydraulic pressure supply part 22 Culture liquid storage part 24 Culture liquid supply pipe 26 Supply amount control part 28 Supply pressure control part 30 Cell Recovery unit 32 Cell storage unit 34 Cell discharge tube 36 Discharge amount control unit 50 Cell transfer unit 52 Cell transfer tube 54 Transfer amount control unit 60 Third hydraulic pressure supply unit 62 Culture solution storage unit 64 Culture solution supply tube 66 Supply amount control unit 68 Supply pressure control unit 100 Cell culture module 101 First cell culture unit 102 Second cell culture unit 110 Second sealing system member 110A Lower lid 110B Upper lid 112 Second internal space 115 Fixing member 120 Second hydraulic pressure supply Unit 122 culture solution storage unit 124 culture solution supply pipe 126 supply amount control unit 128 supply pressure control unit 160 fourth hydraulic pressure supply unit 162 culture solution storage unit 164 culture solution supply tube 66 Supply amount control unit 168 Supply pressure control unit 500 First culture solution circulation system 512 Culture solution storage unit 514 Supply device 516a Pipe 516b Pipe 516c Pipe 518a Circulation amount control unit 518b Circulation amount control unit 600 Second culture solution circulation system 612 Culture solution storage unit 614 Supply device 616a Piping 616b Piping 616c Piping 618a Circulating amount control unit 618b Circulating amount control unit S ₁ First cell culture support S ₁₁ First recess S ₂ Second cell culture support S ₂₂ 2nd Recess in

Claims (7)

A first closed system member for holding a first cell culture carrier made of a porous body provided with a first recess for culturing cells in the first internal space in a sealable manner, and culturing in the first internal space In a state filled with liquid, the first cell culture carrier is held in the first internal space with respect to the second surface facing the first surface provided with the first recess of the first cell culture carrier. A first cell culture section comprising a first water pressure supply section for supplying water pressure;
A second sealing system member for holding a second cell culture carrier comprising a porous body provided with a second recess larger than the first recess for culturing cells in the second internal space so as to be able to be sealed; In a state where the second internal space is filled with a culture solution, a second surface facing the third surface provided with the second concave portion of the second cell culture carrier held in the second internal space. A second cell culture section comprising a second water pressure supply section for supplying a second water pressure to the four surfaces;
The first internal space and the second internal space are connected, and the cells detached from the first recess by the first water pressure are transferred from the first internal space to the second internal space. A cell transport section to perform,
A cell recovery unit connected to the second internal space at a position different from the cell transport unit, and recovering cells detached from the second recess by the second water pressure from the second internal space;
A cell culture module comprising: A first closed system member for holding a first cell culture carrier made of a porous body provided with a first recess for culturing cells in the first internal space in a sealable manner, and culturing in the first internal space A second surface facing the first surface of the first cell culture carrier held in the first inner space and provided with the first recess and the first surface provided with the first recess in a state filled with a liquid. A first water pressure supply unit for supplying a first water pressure to the first cell culture unit,
A second sealing system member for holding a second cell culture carrier comprising a porous body provided with a second recess larger than the first recess for culturing cells in the second internal space so as to be able to be sealed; The second concave portion and the second concave portion of the second cell culture carrier held in the second internal space are provided in a state where the second internal space is filled with a culture solution. A second water culture unit comprising a second water pressure supply unit for supplying a second water pressure to the fourth surface opposite to the three surfaces;
The first internal space and the second internal space are connected, and the cells detached from the first recess by the first water pressure with respect to the second surface of the first cell culture carrier are A cell transport section for transporting from one internal space to the second internal space;
The cell connected to the second internal space at a position different from the cell transfer unit, and the cells detached from the second recess by the second water pressure with respect to the fourth surface of the second cell culture carrier are A cell recovery unit for recovering from the internal space of 2,
A cell culture module comprising:   One or both of the first internal space and the second internal space are filled with a culture solution, and a culture solution circulation system for circulating the filled culture solution is further provided. The cell culture module according to claim 1 or 2.   The connection part connected to the second internal space of the cell transfer part is provided immediately above the second recess of the second cell culture carrier held in the second internal space. The cell culture module according to any one of claims 1 to 3, wherein   The second internal space is newly provided with cell rectification means for guiding the cells transferred from the cell transfer section into the second recesses of the second cell culture carrier. The cell culture module according to any one of claims 1 to 4.   6. The cell culture module according to claim 5, wherein the cell rectifying means is a rectifying plate provided in the second internal space. The size of the space that constitutes the second recess is equal to and larger than the size of the space that constitutes the first recess in all directions, according to any one of claims 1 to 6. Cell culture module.

Images