WO2004005455A1

WO2004005455A1 - Cell culture carrier and cell culture unit

Info

Publication number: WO2004005455A1
Application number: PCT/JP2003/008472
Authority: WO
Inventors: Yoichi Ishikawa
Original assignee: Able Corporation
Priority date: 2002-07-05
Filing date: 2003-07-03
Publication date: 2004-01-15
Also published as: AU2003246257A1

Abstract

A cell culture unit comprising a porous rigid cylindrical cell culture carrier comprising a hard material (excluding hydroxyapatite) and/or a soft material as a carrier material, a cylindrical envelope encircling via an interspace the cylindrical carrier, and culture fluid circulating means for circulating the culture fluid from the interspace via a circumferential wall surface of the cylindrical carrier toward the core portion thereof. The cell culture carrier comprises a carrier material and a cell cure promoter, and is porous, rigid and cylindrical. The cell cure promoter is a polymeric adhesive.

Description

Description Cell culture carrier and cell culture device

The present invention relates to a cell culture carrier and a cell culture device, and more particularly, to a cell culture carrier effective for rapidly, uniformly and densely culturing animal cells in a carrier, a method for producing the same, and a cell culture device. . In addition, cell modules that carry cultured cells, especially animal cells, can be effectively used in many fields such as production of useful substances, safety evaluation of foods and drugs, artificial organs and tissues, and pharmacology. . Background art

Conventionally, as a cell culture reactor, cylindrical large and small nets are arranged in double, and the space surrounded by them is filled with particulate porous cell carriers, and the center from the outer side of the large mesh A reactor has been put into practical use in which a culture solution flows in a reverse radial direction toward the reactor. This reactor is an excellent reactor capable of uniformly supplying a culture solution to cells growing inside the carrier, on the surface of the carrier, or in the space between the carriers, if the particulate carrier is uniformly and closely packed.

However, such a reactor requires a double-arranged cylindrical net to form a space for filling the carrier, and the structure is complicated. It is necessary to pack the carrier uniformly because the culture solution flows.However, the filling operation is difficult because the carrier is particulate, and the carrier is placed inside a cylindrical mesh. Therefore, there was a problem that the carrier could not be observed during the culture. Conventionally, cells have been cultured using the above-mentioned reactors, so only granular carriers are commercially available as carriers, and other forms of carriers are not in circulation. The carrier on which the cells were grown was not sold.

In the cell culture using the above-described reactor, it was difficult to remove the carrier on which the cells had formed from the portion surrounded by the double cylinder. Disclosure of the invention

The present invention has been made in view of the above-mentioned problems of the conventional technology, and does not require a double-placed cylindrical net for accommodating a carrier, does not require an operation of filling the carrier, and provides an animal. It is an object of the present invention to provide a cell culture device capable of culturing cells rapidly, homogeneously and at high density, and observing a carrier during culturing, a cell culture carrier and a cell module on which cells are formed.

The present inventors have conducted intensive studies in order to solve the above problems, and as a result, if a porous carrier having a predetermined shape and properties is formed in a cylindrical block in advance, there is no need to fill a granular carrier, The inventors have found that the problem can be solved, and have completed the present invention.

That is, the first cell culture device of the present invention comprises a porous and cylindrical cell culture carrier containing a hard material (excluding hydroxy and hydroxyapatite) and Z or a soft material as a carrier material. A cylindrical envelope surrounding the cylindrical carrier through a space; and a culture solution distribution means for flowing the culture solution from the space through the outer peripheral wall surface of the cylindrical carrier toward the center. And

In a preferred embodiment of the first cell culture device of the present invention, the hard material is at least one kind of porous or non-porous material selected from the group consisting of metal, glass and ceramics. If the soft material is porous or non-porous Characterized in that it is a polymer material of

Further, another preferred embodiment of the first cell culture device of the present invention is characterized in that the hard material and the Z or soft material are in the form of beads having a particle diameter of 100 to 100 x m.

On the other hand, the cell culture carrier of the present invention contains a carrier material and a cell growth promoting agent, and is characterized by being porous and cylindrical.

Further, in a preferred form of the cell culture carrier of the present invention, the cell growth promoting agent is a polymer-based adhesive, specifically, polyvinyl acetate, polyvinyl alcohol, silicone rubber, natural rubber, and rubber. At least one member selected from the group consisting of loprene rubber, vinyl resin, epoxy resin, phenolic resin, urethane resin, unsaturated polyester, cellulose, protein, starch, sugar, lipid, collagen and derivatives thereof It is characterized by the following. Further, in another preferred form of the cell culture carrier of the present invention, the carrier material is a porous or non-porous hard material and Z or a soft material. In this case, the hard material is metal, glass and At least one kind of porous or non-porous material selected from the group consisting of ceramics, and the soft material is a porous or non-porous polymer material.

Still another preferred embodiment of the carrier for cell culture of the present invention is characterized in that the carrier material is in the form of beads of 100 to 100 μιη.

In this case, it is preferable that the beads of the carrier material are porous and have continuous pores having an opening diameter of 50 μιη or less.

Further, the method for producing a cell culture carrier of the present invention, when producing the cell culture carrier as described above,

Forming a porous cylindrical member using the carrier material,

Next, the cylindrical member is brought into contact with a liquid in which the cell adhesion promoting agent is dissolved or suspended, and then dried. Further, another method for producing the cell culture carrier of the present invention includes the steps of:

The porous or non-porous hard and / or soft material beads are formed into a cylindrical shape by contacting the above-mentioned liquid in which the cell growth agent is dissolved or suspended, and then formed into a cylindrical shape. Drying the body.

On the other hand, the second cell culture device of the present invention is a cell culture device using the cell culture carrier as described above,

This cell culture carrier, a cylindrical envelope surrounding the carrier via a space, and a culture solution flowing means for flowing a culture solution from the space through the outer peripheral wall surface of the carrier toward the center. It is characterized by having.

In a preferred embodiment of the second cell culture device of the present invention, the porosity of the peripheral wall of the carrier is 30 to 85 V o 1 ° / ₀ .

Further, another preferred embodiment of the second cell culture device of the present invention is characterized in that an outer peripheral wall and / or an inner peripheral wall of the carrier is coated with a porous layer having a larger pressure loss than the peripheral wall. I do.

Further, the cell module of the present invention is characterized in that animal cells are formed on the cell culture carrier in the first cell culture device as described above or on the cell culture carrier as described above.

Further, a preferred embodiment of the cell module of the present invention is characterized in that the above-mentioned animal cells are formed using the first cell culture device or the second cell culture device as described above.

In the cell culture apparatus of the present invention, since a cylindrical porous carrier formed integrally is used, there is no need for a double-walled cylindrical net for accommodating a granular carrier as in the related art. ·

In addition, as long as a homogeneous porous carrier is available, the complicated operation of filling the particulate carrier by the user is omitted and the particulate carrier is filled. In this way, it is possible to avoid carrier packing that may occur due to this, and it is possible to achieve uniform and highly reproducible cell culture.

Furthermore, since the porous carrier has an integral structure, the handling of the carrier after cell culture is improved.

Since the cell culture carrier of the present invention has a predetermined cell growth promoting agent added thereto, the growth and growth of the target cells can be further accelerated and accelerated. Since it can function as a binder for the carrier material, it is also effective when imparting a predetermined shape to the cell culture carrier of the present invention.

Further, in the cell culture device of the present invention, since a cylindrical mesh is not required as described above, a transparent cylindrical envelope surrounding the outer peripheral wall of the carrier through a space is arranged. If the culture solution is supplied in a reverse radial direction (radially toward the center), the outer peripheral wall surface of the carrier can be visually recognized, and the culture state can be observed.

Hereinafter, the present invention will be described in detail. In this specification, “%” indicates mass percentage unless otherwise specified.

As described above, the cell culture device of the present invention includes the first cell culture device and the second cell culture device, but has basically the same configuration except that the cell culture carrier used is different.

The cylindrical envelope used in the first and second cell culture devices may be any as long as it can surround or store a cell culture carrier in a cylindrical shape, typically a donut shape, through a space. Examples include a housing, a cylindrical container, and a cylindrical tank. During cell culture, a cell culture solution is filled in the space.

In addition, the culture solution flowing means used in the first and second cell culture devices has a function of flowing the culture solution filled in the space through the peripheral wall of the cylindrical cell culture carrier and into the inside of the cylinder. If you have + minutes, various pumping or Can be exemplified by a suction pump.

In the present invention, the cell culture carrier of the present invention containing a cell adhesion promoting agent is used for the second cell culture device.

The difference between the cell culture carrier of the present invention and the cell culture carrier used in the first cell culture device is, in principle, whether or not it contains a cell growth promoting agent. The carrier will be described.

First, the cell culture carrier used in the first cell culture device (hereinafter, referred to as “first cell culture carrier”) is cylindrical and porous.

Here, the porosity is affected by the type of the cells to be cultured and the culture medium (medium), but it is sufficient if the culture medium can be circulated. 30 to 85 vo 1. /. It is sufficient to have a porosity of.

If the porosity is less than 30%, the carrier volume utilization efficiency is insufficient. If the porosity exceeds 85%, cells may easily flow out or the carrier strength may be insufficient. If the porosity exceeds 85%, the pressure loss will be small, and the culture solution may not flow through the entire carrier but may flow partially. The porosity can be measured by applying a normal bulk density measurement method.

Furthermore, the thickness of the peripheral wall is also affected by the type of cells to be cultured and the type of culture medium, but is preferably 2 cm or less. Dissolved oxygen may be insufficient on the downstream side, specifically, near the central cavity of the cylindrical carrier, which is not preferable. The carrier material is roughly classified into a hard material and a soft material. In the present invention, both can be used, and both can be used in combination. Such hard materials refer to various metals, inorganic materials such as glass and ceramics, and soft materials refer to various resins, cellulose, elastomers, and other materials. Means a polymer material such as However, hydroxyapatite shall be excluded from hard materials (inorganic materials).

Such a hard material and a soft material can be used regardless of whether they are porous or non-porous, and a mixture of a porous material and a non-porous material can be used. .

A method for producing the first cell culture carrier using the above-described hard material or soft material will be described.

First, when using an inorganic hard material, for example, glass or ceramic, a raw material powder or the like may be molded and fired to form a cylindrical porous glass or porous ceramic by a conventionally known method.

In the case of a polymer material, a cylindrical porous resin molded body may be molded by a so-called foam molding method using various foaming agents, for example, a cylindrical foamed polyurethane resin molded body may be manufactured. In addition, a porous resin molded article can be obtained by a pressure blowing method of an inert gas such as nitrogen gas.

In addition, a non-woven fabric (strip) such as a polyester non-woven fabric may be filled in a mold having an appropriate cylindrical cavity, and a binder may be added to the mold to bond the non-woven fabric. Alternatively, a cylindrical body may be created.

In addition, it is a matter of course that the carrier may be formed by cutting out a relatively large porous material into a desired shape.

On the other hand, as another manufacturing method, there is a method in which beads (particles) of a hard material or a soft material are bonded or bound by an appropriate method to form a porous cylindrical body.

For example, in the case of metal, glass, and ceramics, the metal beads, glass beads, and ceramic beads are assembled into a cylindrical shape, heated, and the partially melted beads are bound together to form a porous cylinder. Shape the body Can be achieved.

The particle size of the beads at this time is not particularly limited, but may be 100 to 10Ομπι.

A porous cylinder can also be obtained by binding metal particles, glass particles, ceramic particles, resin particles, rubber particles, and the like with an appropriate binder.

The beads used in such a production method may be porous or non-porous, but porous beads may be more preferable in consideration of the flowability of the culture solution. Examples of porous ceramics include silica gel, bentonite, alumina, silica, zeolite, and magnesia.

When porous beads are used, it is preferable to select those having continuous holes with an opening diameter of 50 m or less in order to ensure the flowability of the culture solution. In a pore having an opening diameter of 50 μm or less, it is easy to secure a flow path of a culture solution because cells hardly proliferate inside the pore.

Next, the cell culture carrier of the present invention (hereinafter referred to as “second cell culture carrier”) will be described.

This second cell culture carrier has basically the same configuration as the above-described first cell culture carrier except that it contains a cell growth promoting agent. However, the carrier material of the second cell culture carrier also includes hydroxyapatite. Hydroxyapatite is also used in artificial bones and dental roots, and has the property of easily adhering cells.

Here, the cell adhesion promoting agent is not particularly limited as long as it promotes cell adhesion, and is not particularly limited. Examples thereof include a so-called polymer adhesive. That is, high molecular compounds having adhesive properties, for example, poly (vinyl acetate), poly (vinyl alcohol), poly (vinyl chloride), cyanoacrylate. Thermoplastic resins such as polystyrene, polyamide and unsaturated polyester; synthetic resins such as epoxy resin, phenolic resin and urethane resin; rubbers such as natural rubber, nitrile rubber and silicone rubber; cellulose; Mention may be made of starch, sugar, lipid or collagen, and any mixtures thereof.

In addition, as such a polymer compound, not only the above-mentioned compounds and materials themselves, but also various derivatives such as ion-exchange derivatives can be used, and these derivatives can be used in any combination of at least one kind. it can.

These adhesive polymer compounds are usually dissolved or suspended in an appropriate solvent or in the form of an emulsion, for example, a methanol solution of polyvinyl acetate, an aqueous solution of polyvinyl alcohol, or a rubber paste. It is used in the form of aqueous solutions of emulsion rubber, cellulose, starch, sugar and collagen of black rubber, vinyl resin, epoxy resin, fuanol resin, urethane resin and unsaturated polyester resin.

The cell growth promoting agent as described above can be contained in the cell culture medium by a coating method-dipping method.

Typically, a solution or an emulsion of the cell growth promoter is applied to the surface of the first cell culture carrier, or the first cell culture carrier is immersed in the solution or the emulsion of the cell growth promoter. Further, it may be dried by air drying or hot air drying. In addition, such application and immersion may be repeated not only once but also several times to ensure the content of the cell growth promoter.

On the other hand, since the cell growth promoting agent has adhesiveness and binding properties, the second cell culture carrier can also be produced by binding the beads of the above carrier material with the cell growth promoting agent.

Specifically, a carrier material bead is filled into a mold with cylindrical cavity. After that, a solution of the cell growth promoting agent may be poured, or the cell growth promoting solution may be poured into a mold first, filled with carrier material beads, and then dried. By such drying, the solvent contained in the cell adhesion promoter solution or the like is released, so that a porous carrier as a whole can be obtained even if the carrier material beads are non-porous.

Further, in the present invention, in the first and second cell culture carriers, it is possible to coat the outer peripheral wall surface and / or the inner peripheral wall surface with a porous layer having a larger pressure loss than the carrier itself. The effect of uniformly flowing the culture solution through the carrier can be further improved.

Such a porous layer having a large pressure loss may be formed by forming a porous pipe having a large pressure loss in advance and bringing a carrier into close contact with the porous pipe, or forming a porous pipe concentric with the carrier in the envelope. It can also be formed by arranging pipes.

The cell module of the present invention is obtained by growing animal cells on the first cell culture carrier or the second cell culture carrier described above. Typically, these carriers are cultured in the cell culture device of the present invention. It was done.

The first cell culture carrier and the second cell culture carrier, particularly the second cell culture carrier, are excellent in the formation and cultivation of animal cells, and the uniformity of porosity, Excellent culture uniformity due to isotropic properties, and excellent handling properties during transport / attachment / detachment due to rigidity. BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 is a cross-sectional view showing one example of the cell culture reactor of the present invention, and FIG. 2 is a configuration diagram showing one example of the cell culture system of the present invention. -Best mode for carrying out the invention Hereinafter, the present invention will be described in more detail with reference to some examples, but the present invention is not limited to these examples.

[Cell culture device]

(Example 1)

FIG. 1 is a cross-sectional view showing one example of the cell culture reactor of the present invention.

In this cell culture reactor 31, the bottom plate 1 is formed integrally with the central column 2. Further, an integrally formed cylindrical porous carrier block 4 and a glass cylinder 5 are disposed in contact with the elastic sheet 3 disposed on the upper surface of the bottom plate 1, and the elastic sheet 6 and the top plate 7 are disposed in contact with the upper end thereof. The bottom plate 1, the top plate 7, the elastic sheets 3 and 6 and the glass cylinder 5 form a substantially cylindrical envelope. Then, the female screw 9 of the culture solution outlet nozzle 8 is screwed into the male screw 10 formed at the upper end of the column 2 to form the bottom plate 1, the elastic sheets 3 and 6, the porous carrier 4, the glass cylinder 5, and the culture solution. Outlet nozzle 8 is fixed integrally.

A culture solution supply nozzle 11 is fixed to the top plate 7 by being sealed with an O-ring 12. The O-ring 13 prevents the culture solution from leaking from the screw portion between the top plate 7 and the culture solution outlet nozzle 8. The glass cylinder 5 and the outer peripheral wall surface of the porous carrier 4 are separated from each other, and a space 14 is formed and communicates with the culture solution supply nozzle 11.

Next, the operation of the cell culture reactor will be described.

When the culture solution is supplied from the culture solution supply nozzle 11 by a pump, the culture solution fills the space 14 and flows in the direction of arrow 23 from the outer periphery of the cylindrical carrier 4 in a reverse radial direction to form a hollow portion 2 1 at the center. To reach. A part of the column 2 is thin, from which the culture solution is introduced into the hollow portion 21 in the center of the column 2 through a plurality of lateral holes 22 and discharged from the culture solution outlet nozzle 8.

As described above, the porous carrier 4 may be made of various hard materials or soft materials. What uses a material as a carrier material can be used.

Further, by coating or containing the cell growth promoter of the present invention on the porous carrier 4 (not shown), the cell culture carrier of the present invention can be obtained. The cell culture reactor using the cell culture carrier containing the agent corresponds to the second cell culture device of the present invention.

It is desirable to select an appropriate material for the porous carrier 4 in consideration of the type of the cell, the purpose of the culture, the culture conditions, and the like.

[Cell culture system]

(Example 2)

FIG. 2 is a configuration diagram showing an example of a culture system using the cell culture reactor of the present invention.

In this culture system, a culture medium 33 in a medium adjustment tank 32 is circulated to a reactor 31 in FIG. 1 by a pump 34. In the medium 33, the pH, dissolved oxygen, dissolved carbon dioxide, and composition of the culture solution are controlled by ordinary methods. An example of culture using this cell culture device is described below.

[Culture example] ''

When cells are put into the medium in the medium adjusting tank 32 and sent into the reactor 31 by the pump 34, the cells are sent to the porous carrier 4 and seeded. At that time, it is necessary to appropriately control the flow rate so that the seed is uniformly seeded on the entire porous carrier. After the cells are seeded in the carrier, culture is performed at a different flow rate if necessary. When CHO-kl (Chinese Hamster O vary) was cultured using a porous glass with a porosity of 80%, a diameter of 3 cm, and a hollow diameter of 6 mm as a carrier, the ratio was 1 per m1 of carrier volume. 0 were grown in ^eight of the cell density. · ·

In addition to the above procedure, the cell seeding method includes a method of immersing the porous carrier of the present invention in a cell suspension, and a method of forming a porous carrier into a cell suspension in a closed container. It is possible to adopt a method of immersing and applying a negative pressure to evacuate the air in the carrier and distribute the cells in the carrier.

When the cells are seeded on the carrier, the cells are allowed to stand and adhere, and the post-culture solution is allowed to flow, so that the cells are prevented from flowing out of the carrier.

In the present invention, it is desirable to provide a part that generates a pressure loss in the carrier in order to allow the culture solution to flow uniformly in the porous carrier.

That is, in the cylindrical porous carrier of the present invention, when the culture solution flows from the outer peripheral wall to the center, the flow rate increases near the center and the pressure loss at the center increases, so that the culture solution easily flows uniformly. Although it has a structure, it is preferable to arrange a porous body having a larger pressure loss than the material constituting the porous carrier on the outer peripheral wall or the inner peripheral wall of the porous carrier in order to flow more uniformly.

Here, the porous body arranged on the outer peripheral wall or the inner peripheral wall may be in close contact with the porous carrier, but may be in the flow path through which the culture solution flows even if it does not adhere. The column 2 of the above embodiment may be formed of a dense porous body having a hollow portion. In this case, the side hole 22 is unnecessary.

Further, a liquid dispersion layer may be provided in close contact with the outer peripheral wall of the porous carrier or on the outer edge. As the liquid dispersion layer, a porous body or a cylinder having fine holes can be used.

[Cell culture carrier]

(Example 3)

A vertical hole is formed along the center axis of the porous glass cylinder to form a cylindrical porous carrier, and a porous body having the same shape as the porous carrier 4 of the cell culture reactor shown in Fig. 1 is obtained. Created. A 20% methanol solution of polyacetate butyl was applied to the obtained porous body with a brush and air-dried to obtain a cell culture carrier of this example.

The obtained cell culture carrier is installed in the cell culture reactor shown in Fig. 1 and cultured. When the culture solution was supplied from the nutrient solution supply nozzle 11, the culture solution filled the space 14 and directed from the outer wall surface of the cylindrical porous carrier to the inner wall surface in the direction of arrow 23 (reverse radius direction). It was confirmed that the water flowed into the column 2 via the lateral hole 22 and reached the hollow portion of the column 2, and was further discharged from the culture solution outlet nozzle 8.

If such a cell culture carrier is mounted on a cell culture device, cells to be cultured are inoculated, and if the supply of the culture solution, the flow through the porous carrier, and the discharge can be continuously performed, the cells can be cultured in a large amount. Can be.

(Example 4)

The established hepatocytes HepG2 were put into a medium adjusting tank, and the obtained cell-containing medium was passed through the cell culture device equipped with the cell culture carrier of Example 3 to seed animal cells.

After the turbidity of the culture medium adjusting tank disappeared, the flow rate of the culture solution was reduced to half and the flow of the culture solution was continued. After that, while controlling the flow rate so that the dissolved oxygen can be maintained at the outlet of the carrier, the cell density can be adjusted to 10 per 1 m 1 of the carrier volume based on the consumption rate of darcos cultured while continuously changing the culture medium. ^{When the} cell culture carrier reached ⁸ cells, the cell culture carrier was taken out. As a result, a cell module in which animal cells were grown uniformly and at high density was formed.

As described above, the present invention has been described in detail with reference to some examples. However, the present invention is not limited to these examples, and various modifications can be made within the scope of the present invention.

For example, the circular carrier may be formed by molding a soft material such as expanded polyurethane, cellulose, or by cutting or punching out a large block (block). May be.

If a carrier made of such a soft material is used and appropriately crushed and stored, the elastic sheets 3 and 6 in FIG. 1 can be omitted. Industrial applicability

As described above, according to the present invention, a porous carrier having a predetermined shape and properties is formed into a block shape in advance. No mesh is required, no operation for filling the carrier is required, animal cells can be cultured quickly, uniformly and at high density, and a cell culture device, a cell culture carrier and cells that can observe the carrier during culture are formed. The cell module can be provided.

For example, the cell culture device of the present invention enables co-culture of buoyant or adherent cells / heterologous cells.

In particular, hepatocytes are responsible for many metabolic functions in vivo, and the cell module constructed by culturing hepatocytes at high density according to the present invention can be used for substance production, food and drug safety evaluation, and liver function evaluation. In addition to pharmacological evaluation, it is also effective as a device for replicating viruses by introducing a virus such as hepatitis or its gene into the cells of the module.

Claims

The scope of the claims

1. A porous and cylindrical cell culture carrier containing a hard material (except for hydroxyapatite) and Z or a soft material as a carrier material, and a cylinder surrounding the cylindrical carrier via a space A cell culture device, comprising: a shape envelope; and a culture solution flowing means for flowing the culture solution from the space through the outer peripheral wall surface of the cylindrical carrier toward the center.

2. The cell culture device according to claim 1, wherein the hard material is at least one kind of porous or non-porous material selected from the group consisting of metal, glass, and ceramics.

3. The cell culture device according to claim 1, wherein the soft material is a porous or non-porous polymer material.

4. The cell culture device according to any one of claims 1 to 3, wherein the hard material and / or the soft material are in the form of beads having a particle size of 100 to 100 ιη. .

5. A cell culture carrier comprising a carrier material and a cell growth promoting agent, characterized by being porous and cylindrical.

6. The cell culture carrier according to claim 5, wherein the cell growth promoting agent is a polymer adhesive.

7. The above cell adhesion promoting agent is polyacetate bur, polybutyl alcohol. Silicone rubber, natural rubber, chloroprene rubber, bur resin, epoxy resin, phenol resin, urethane resin, unsaturated polyester, cellulose, protein, The cell culture carrier according to claim 5, wherein the carrier is at least one member selected from the group consisting of starch, sugar, lipid, collagen, and derivatives thereof.

8. The carrier material is a porous or non-porous hard material and Ζ or 8. The carrier for cell culture according to claim 5, wherein the carrier is a soft material.

9. The carrier for cell culture according to claim 8, wherein the hard material is at least one kind of porous or non-porous material selected from the group consisting of metal, glass and ceramics.

10. The cell culture carrier according to claim 8, wherein the soft material is a porous or nonporous polymer material.

11. The cell culture carrier according to any one of claims 5 to 10, wherein the carrier material is in the form of beads of 100 to 100 // πι.

12. The carrier for cell culture according to claim 11, wherein the beads of the carrier material are porous and have continuous holes having an opening diameter of 50 µm or less.

13. In producing the carrier for cell culture according to any one of claims 5 to 12,

Forming a porous cylindrical member using the carrier material,

Subsequently, the cylindrical member is brought into contact with a liquid in which the above-mentioned cell growth promoting agent is dissolved or suspended, and thereafter, dried, and thereafter dried.

14. In producing the cell culture carrier according to claim 11 or 12, a porous or non-porous hard material bead and / or a soft material bead are dissolved or suspended in the cell growth agent. Into a cylindrical shape by contact with the liquid

Thereafter, the cylindrical molded body is dried. A method for producing a cell culture carrier.

15.A cell culture apparatus using the cell culture carrier according to any one of claims 5 to 12,

The cell culture carrier and a cylindrical member surrounding the carrier via a space. A cell culture device, comprising: a rope; and a culture solution flowing means for flowing the culture solution from the space through the outer peripheral wall surface of the carrier toward the center.

16. The cell culture device according to claim 15, wherein the porosity of the peripheral wall of the carrier is 30 to 85 vol%.

17. The cell culture according to claim 15, wherein the outer peripheral wall and the inner or inner peripheral wall of the carrier are coated with a porous layer having a larger pressure loss than the peripheral wall. apparatus.

18. A cell module comprising a cell culture carrier according to any one of claims 1 to 4, wherein animal cells are formed on the cell culture carrier.

19. The cell modulo according to claim 18, wherein the animal cells are formed using the cell culture device according to any one of claims 1 to 4.

20. A cell module obtained by growing animal cells on the cell culture carrier according to any one of claims 5 to 12.

21. The cell module according to claim 20, wherein the animal cells are formed using the cell culture device according to any one of claims 15 to 17.