Cell scaffold material, cell culture substrate, and method for culturing t cells

Abstract

Provided is a cell scaffold material that can bring about good maintenance of memory T cells and good T cell proliferation.　A cell scaffold material according to the present invention is used for culturing a cell population containing T cells and contains a peptide-containing resin (A) having a synthetic resin part and a peptide part. The peptide part has a first peptide part and a second peptide part. The first peptide part and the second peptide part have different amino acid sequences. The first peptide part can bind to a first integrin, and the second peptide part can bind to a second integrin. The dispersive component γd of the surface free energy of the cell scaffold material is at least 24.5 mJ / m2 and not more than 45.0 mJ / m2, and the dipole component γp of the surface free energy of the cell scaffold material is at least 1.0 mJ / m2 and not more than 20.0 mJ / m2.

Description

Cell scaffold materials, cell culture substrates, and methods for culturing T cells

[0001] This invention relates to a cell scaffold material. Furthermore, this invention relates to a cell culture substrate comprising the above-mentioned cell scaffold material. Moreover, this invention relates to a method for culturing T cells using the above-mentioned cell scaffold material.

[0002] Surgical intervention, chemotherapy, and radiation therapy are widely used as treatments for cancer. In recent years, immunotherapy has been attracting attention as a fourth cancer treatment method. Known immunotherapies include methods that administer PD-1 inhibitors, which have an inhibitory effect on PD-1, an immune checkpoint molecule expressed on immune cells, and methods that administer T cells with enhanced antitumor effects (e.g., CAR-T therapy and TCR-T therapy).

[0003] Immunotherapy such as CAR-T therapy and TCR-T therapy requires culturing the desired T cells in vitro.

[0004] For example, Patent Document 1 below discloses a method for producing a cell population into which a desired gene has been introduced, comprising the following steps (1) and (2): (1) A step of culturing a cell population containing T cells and / or T cell precursor cells in a container containing fibronectin or a fragment thereof and a CD3 ligand. (2) A step of adding a vector containing the desired gene to the container of step (1).

[0005] WO2009 / 119793A1

[0006] In immunotherapy, it is expected that administering memory T cells, which are in a more undifferentiated state, to the body will yield higher therapeutic effects than administering terminally differentiated effector T cells.

[0007] Incidentally, conventionally, cell adhesion proteins such as fibronectin, as described in Patent Document 1, have been widely used as cell scaffold materials for culturing T cells. Cell scaffold materials containing synthetic resins are also known. However, in conventional T cell culture methods, T cell differentiation tends to progress during culture. In particular, when conventional cell scaffold materials containing synthetic resins are used, T cell differentiation tends to progress during culture. Therefore, with conventional T cell culture methods, it can be difficult to maintain memory T cells well while promoting good T cell proliferation.

[0008] The object of the present invention is to provide a cell scaffold material that can maintain memory T cells well and promote the proliferation of T cells well. The present invention also aims to provide a cell culture substrate comprising the above cell scaffold material. Furthermore, the present invention aims to provide a method for culturing T cells using the above cell scaffold material.

[0009] This specification discloses the following cell scaffold materials, cell culture substrates, and methods for culturing T cells.

[0010] Item 1. A cell scaffold material used for culturing a cell population including T cells, comprising a peptide-containing resin (A) having a synthetic resin portion and a peptide portion, wherein the peptide portion comprises a first peptide portion and a second peptide portion, the first peptide portion and the second peptide portion having different amino acid sequences, the first peptide portion being capable of binding to a first integrin, the second peptide portion being capable of binding to a second integrin, and the dispersion component γ of the surface free energy of the cell scaffold material. ｄ However, 24.5 mJ / m ２ 45.0mJ / m or more ２ The dipole component γ of the surface free energy of the cell scaffold material is as follows: ｐ However, 1.0 mJ / m ２ 20.0mJ / m or more ２ The following are cell scaffold materials.

[0011] Item 2. The cell scaffold material according to Item 1, wherein the first integrin is VLA-5 and the second integrin is VLA-4.

[0012] Item 3. The cell scaffold material according to item 1 or 2, wherein the first peptide portion has an RGD sequence and the second peptide portion has an LDV sequence.

[0013] Item 4. The cell scaffold material according to any one of items 1 to 3, wherein the first peptide portion has a c-RGDfK sequence.

[0014] Item 5. The cell scaffold material according to any one of items 1 to 4, wherein the second peptide portion has an EILDV sequence.

[0015] Item 6. The cell scaffold material according to any one of items 1 to 5, wherein the peptide-containing resin (A) has a total content P1 of the first peptide portion and a total content P2 of the second peptide portion of 0.02 mol% or more and 4.0 mol% or less.

[0016] Item 7. The cell scaffold material according to any one of items 1 to 6, wherein the peptide-containing resin (A) has a content P1 of the first peptide portion of 0.01 mol% or more and a content P2 of the second peptide portion of 0.01 mol% or more and a content P2 of 0.01 mol% or more and a content P2 of 2.0 mol% or less.

[0017] Item 8. The cell scaffold material according to any one of items 1 to 7, wherein the synthetic resin portion is a polyvinyl acetal resin portion or a (meth)acrylic polymer portion.

[0018] Item 9. The cell scaffold material according to any one of items 1 to 8, wherein the synthetic resin portion is a polyvinyl acetal resin portion.

[0019] Item 10. The cell scaffold material according to any one of items 1 to 9, wherein the peptide-containing resin (A) has a linker portion, and the synthetic resin portion and the peptide portion are bonded via the linker portion.

[0020] Item 11. The cell scaffold material according to item 10, wherein the linker portion has structural units derived from polymerizable monomers.

[0021] Item 12. The cell scaffold material according to item 11, wherein the polymerizable monomer is a (meth)acrylate compound.

[0022] Item 13. The cell scaffold material according to Item 11 or 12, wherein the polymerizable monomer contains a compound having a carboxyl group.

[0023] Item 14. A cell scaffold material used for culturing a cell population containing T cells, comprising a peptide-containing resin (1) having a synthetic resin part and a first peptide part, and a peptide-containing resin (2) having a synthetic resin part and a second peptide part, wherein the first peptide part and the second peptide part have different amino acid sequences, the first peptide part is capable of binding to a first integrin, the second peptide part is capable of binding to a second integrin, and the dispersive component γ of the surface free energy of the cell scaffold material ｄ is 24.5 mJ / m ２ or more and 45.0 mJ / m ２ or less, and the dipole component γ of the surface free energy of the cell scaffold material ｐ is 1.0 mJ / m ２ or more and 20.0 mJ / m ２ or less. The cell scaffold material.

[0024] Item 15. The cell scaffold material according to Item 14, wherein the first integrin is VLA-5 and the second integrin is VLA-4.

[0025] Item 16. The cell scaffold material according to Item 14 or 15, wherein the first peptide part has an RGD sequence and the second peptide part has an LDV sequence.

[0026] Item 17. The cell scaffold material according to any one of Items 14 to 16, wherein the first peptide part has a c-RGDfK sequence.

[0027] Item 18. The cell scaffold material according to any one of Items 14 to 17, wherein the second peptide part has an EILDV sequence.

[0028] Item 19. The cell scaffold material according to any one of Items 14 to 18, wherein the total of the content ratio P1 of the first peptide part in the peptide-containing resin (1) and the content ratio P2 of the second peptide part in the peptide-containing resin (2) is 0.02 mol% or more and 4.0 mol% or less.

[0029] Item 20. The cell scaffold material according to any one of items 14 to 19, wherein the content P1 of the first peptide portion in the peptide-containing resin (1) is 0.01 mol% or more and 2.0 mol% or less, and the content P2 of the second peptide portion in the peptide-containing resin (2) is 0.01 mol% or more and 2.0 mol% or less.

[0030] Item 21. The cell scaffold material according to any one of items 14 to 20, wherein the synthetic resin portion in the peptide-containing resin (1) is a polyvinyl acetal resin portion or a (meth)acrylic polymer portion, and the synthetic resin portion in the peptide-containing resin (2) is a polyvinyl acetal resin portion or a (meth)acrylic polymer portion.

[0031] Item 22. The cell scaffold material according to any one of items 14 to 21, wherein the synthetic resin portion in the peptide-containing resin (1) is a polyvinyl acetal resin portion, and the synthetic resin portion in the peptide-containing resin (2) is a polyvinyl acetal resin portion.

[0032] Item 23. The cell scaffold material according to any one of items 14 to 22, wherein the peptide-containing resin (1) has a linker portion, and in the peptide-containing resin (1), the synthetic resin portion and the first peptide portion are linked via the linker portion, and the peptide-containing resin (2) has a linker portion, and in the peptide-containing resin (2), the synthetic resin portion and the second peptide portion are linked via the linker portion.

[0033] Item 24. The cell scaffold material according to item 23, wherein the linker portion in the peptide-containing resin (1) has structural units derived from polymerizable monomers, and the linker portion in the peptide-containing resin (2) has structural units derived from polymerizable monomers.

[0034] Item 25. The cell scaffold material according to item 23 or 24, wherein the linker portion in the peptide-containing resin (1) has structural units derived from a (meth)acrylate compound, and the linker portion in the peptide-containing resin (2) has structural units derived from a (meth)acrylate compound.

[0035] Item 26. The cell scaffold material according to any one of items 23 to 25, wherein the linker portion in the peptide-containing resin (1) has structural units derived from a compound having a carboxyl group, and the linker portion in the peptide-containing resin (2) has structural units derived from a compound having a carboxyl group.

[0036] Item 27. The cell scaffold material according to any one of items 1 to 26, wherein the T cells include CD8+ T cells.

[0037] Item 28. A cell culture substrate comprising a substrate body and a cell scaffold material disposed on a first surface of the substrate body, wherein the cell scaffold material is the cell scaffold material described in any one of items 1 to 27.

[0038] Item 29. The cell culture substrate according to item 28, wherein the substrate body is a container.

[0039] Item 30. A method for culturing T cells, comprising a culture step of culturing a cell population including T cells on the surface of a cell scaffold material described in any one of items 1 to 27.

[0040] The cell scaffold material according to the present invention is a cell scaffold material used for culturing a cell population including T cells. The cell scaffold material according to the present invention comprises a peptide-containing resin (A) having a synthetic resin portion and a peptide portion, wherein the peptide portion has a first peptide portion and a second peptide portion. The first peptide portion and the second peptide portion have different amino acid sequences. In the cell scaffold material according to the present invention, the first peptide portion is capable of binding to a first integrin, and the second peptide portion is capable of binding to a second integrin. In the cell scaffold material according to the present invention, the dispersion component γ of the surface free energy of the cell scaffold material ｄ However, 24.5 mJ / m ２ 45.0mJ / m or more ２ The dipole component γ of the surface free energy of the cell scaffold material is as follows: ｐ However, 1.0 mJ / m ２ 20.0mJ / m or more ２The following is true. The cell scaffold material according to the present invention has the above configuration, so that memory T cells can be well maintained and T cells can be well proliferated.

[0041] The cell scaffold material according to the present invention is a cell scaffold material used for culturing a cell population including T cells. The cell scaffold material according to the present invention comprises a peptide-containing resin (1) having a synthetic resin portion and a first peptide portion, and a peptide-containing resin (2) having a synthetic resin portion and a second peptide portion, wherein the first peptide portion and the second peptide portion have different amino acid sequences. In the cell scaffold material according to the present invention, the first peptide portion is capable of binding to a first integrin, and the second peptide portion is capable of binding to a second integrin. In the cell scaffold material according to the present invention, the dispersion component γ of the surface free energy of the cell scaffold material ｄ However, 24.5 mJ / m ２ 45.0mJ / m or more ２ The dipole component γ of the surface free energy of the cell scaffold material is as follows: ｐ However, 1.0 mJ / m ２ 20.0mJ / m or more ２ The following is true. The cell scaffold material according to the present invention has the above configuration, so that memory T cells can be well maintained and T cells can be well proliferated.

[0042] Figure 1 is a schematic diagram showing an example of a peptide-containing resin (A) that can be used in the present invention. Figure 2(a) is a schematic diagram showing an example of a peptide-containing resin (1) that can be used in the present invention, and Figure 2(b) is a schematic diagram showing an example of a peptide-containing resin (2) that can be used in the present invention. Figure 3 is a schematic cross-sectional view showing a cell culture substrate according to one embodiment of the present invention.

[0043] The details of the present invention will be described below.

[0044] [Cell scaffold material] The cell scaffold material (A) according to the present invention is a cell scaffold material used for culturing a cell population including T cells. The cell scaffold material (A) according to the present invention comprises a peptide-containing resin (A) having a synthetic resin portion and a peptide portion, wherein the peptide portion has a first peptide portion and a second peptide portion. The first peptide portion and the second peptide portion have different amino acid sequences. In the cell scaffold material (A) according to the present invention, the first peptide portion is capable of binding to a first integrin, and the second peptide portion is capable of binding to a second integrin. In the cell scaffold material (A) according to the present invention, the dispersion component γ of the surface free energy of the cell scaffold material (A) ｄ However, 24.5 mJ / m ２ 45.0mJ / m or more ２ The dipole component γ of the surface free energy of the cell scaffold material (A) is as follows: ｐ However, 1.0 mJ / m ２ 20.0mJ / m or more ２ The following applies:

[0045] The cell scaffold material (B) according to the present invention is a cell scaffold material used for culturing a cell population including T cells. The cell scaffold material (B) according to the present invention comprises a peptide-containing resin (1) having a synthetic resin portion and a first peptide portion, and a peptide-containing resin (2) having a synthetic resin portion and a second peptide portion, wherein the first peptide portion and the second peptide portion have different amino acid sequences. In the cell scaffold material (B) according to the present invention, the first peptide portion is capable of binding to a first integrin, and the second peptide portion is capable of binding to a second integrin. In the cell scaffold material (B) according to the present invention, the dispersion component γ of the surface free energy of the cell scaffold material (B) ｄ However, 24.5 mJ / m ２ 45.0mJ / m or more ２ The dipole component γ of the surface free energy of the cell scaffold material (B) is as follows: ｐ However, 1.0 mJ / m ２ 20.0mJ / m or more ２ The following applies:

[0046] Hereinafter, in this specification, cell scaffold material (A) and cell scaffold material (B) may be collectively referred to as cell scaffold material.

[0047] The cell scaffold material according to the present invention has the above-described configuration, and therefore can effectively maintain memory T cells and effectively promote T cell proliferation. Despite using components (resins) that are not cell adhesion proteins, the cell scaffold material according to the present invention can effectively maintain memory T cells and effectively promote T cell proliferation.

[0048] The inventors have discovered that the maintenance performance of memory T cells and the proliferation performance of T cells change depending on the type of components contained in the cell scaffold material. Furthermore, the inventors have found that by using a cell scaffold material that contains a peptide-containing resin having a specific amino acid sequence and has a specific surface free energy, it is possible to maintain memory T cells well and to promote the proliferation of T cells well.

[0049] The cell scaffold material of the present invention allows for the efficient proliferation of T cells while maintaining memory T cells effectively, thus enabling the simple acquisition of cell populations useful for cell therapies such as immunotherapy.

[0050] From the viewpoint of maintaining memory T cells well and promoting T cell proliferation, the dispersion component γ of the surface free energy of the cell scaffold material is important. ｄ It is 24.5 mJ / m ２ 45.0mJ / m or more ２ The following applies:

[0051] Dispersion component γ of the surface free energy of the above cell scaffold material ｄ Preferably 28.0 mJ / m ２ More preferably, 30.0 mJ / m ２ More preferably, 32.5 mJ / m ２ Preferably, the above is 38.0 mJ / m ２ More preferably, 36.0 mJ / m ２ The above is the dispersion component γ. ｄWhen the above lower limit and upper limit are met, memory T cells can be maintained more effectively, and T cells can be proliferated more effectively. Also, the above dispersed component γ ｄ When the values ​​are above the lower limit and below the upper limit, the ability of cells to adhere to the cell scaffold material can be improved.

[0052] From the viewpoint of maintaining memory T cells well and promoting T cell proliferation, the dipole component γ of the surface free energy of the cell scaffold material is important. ｐ This is 1.0 mJ / m ２ 20.0mJ / m or more ２ The following applies:

[0053] The dipole component γ of the surface free energy of the above cell scaffold material ｐ Preferably 2.5 mJ / m ２ Preferably, the above is 10.0 mJ / m ２ More preferably, 5.0 mJ / m ２ The following is the dipole component γ. ｐ When the above lower limit and upper limit are met, memory T cells can be maintained more effectively, and T cells can be proliferated more effectively. Also, the above dipole component γ ｐ When the values ​​are above the lower limit and below the upper limit, the ability of cells to adhere to the cell scaffold material can be improved.

[0054] The dispersion component γ of the above surface free energy ｄ and dipole component γ ｐ This is calculated using the Kaelble-Uy theoretical formula. The Kaelble-Uy theoretical formula is as shown in equation (1) below, where the total surface free energy γ is equal to the dispersion component γ ｄ and dipole component γ ｐ This is a theoretical formula based on the assumption that it will be the sum of [two factors].

[0055]

[0056] Furthermore, in the Kaelble-Uy theoretical formula, the surface free energy of the liquid is γ ｌ (mJ / m ２ ) and the surface free energy of the solid is γ ｓ (mJ / m２ If we assume that the contact angle is θ (°), then the following equation (2) holds true.

[0057]

[0058] Therefore, the surface free energy γ of the liquid ｌ Using two known liquids, the contact angle θ with respect to a resin film (membrane cell scaffold material) formed using cell scaffold material was measured, and γ ｓ ｄ and γ ｓ ｐ By solving the simultaneous equations, the dispersion component γ of the surface free energy of the cell scaffold material can be obtained. ｄ and dipole component γ ｐ It is possible to find this.

[0059] In this invention, the above surface free energy γ ｌ As two known types of liquids, pure water and diiodomethane are used.

[0060] The above contact angle θ is measured using a contact angle meter (for example, "DMo-701" manufactured by Kyowa Interface Chemical Co., Ltd.) as follows.

[0061] 1 μL of pure water or diiodomethane is dropped onto the surface of a resin film formed using a cell scaffold material. The angle between the pure water and the resin film 10 seconds after dropping is defined as the contact angle θ with respect to the pure water. Similarly, the angle between the diiodomethane and the resin film 10 seconds after dropping is defined as the contact angle θ with respect to the diiodomethane.

[0062] The above variance component γ ｄ Methods to increase the amount of nonpolar functional groups in the peptide-containing resin include increasing the amount of nonpolar functional groups and introducing functional groups having a cyclic structure in the peptide-containing resin. ｄ Methods to reduce the dipole component γ include reducing the amount of butyl groups, which are nonpolar functional groups, in the peptide-containing resin. ｐ Methods to increase the dipole component γ include increasing the amount of polar functional groups in the peptide-containing resin, and introducing functional groups containing ether structures into the peptide-containing resin.ｐ Methods to reduce this include increasing the amount of butyl groups, which are nonpolar functional groups, in the peptide-containing resin.

[0063] (Peptide-conjugated resin) The cell scaffold material (A) includes a peptide-conjugated resin (A) having a synthetic resin portion and a peptide portion. The peptide-conjugated resin (A) is a synthetic resin to which peptides are bound. The peptide-conjugated resin (A) has a synthetic resin portion and a peptide portion. The peptide portion has a first peptide portion and a second peptide portion. Therefore, the peptide-conjugated resin (A) has a synthetic resin portion, a first peptide portion and a second peptide portion. The peptide-conjugated resin (A) preferably has a linker portion. In the peptide-conjugated resin (A), it is preferable that the synthetic resin portion and the peptide portion are bound via the linker portion. Only one type of peptide-conjugated resin (A) may be used, or two or more types may be used in combination.

[0064] The cell scaffold material (B) above includes a peptide-containing resin (1) having a synthetic resin portion and a first peptide portion. The cell scaffold material (B) above includes a peptide-containing resin (2) having a synthetic resin portion and a second peptide portion. The peptide-containing resins (1) and (2) above are synthetic resins to which peptides are bound. The peptide-containing resin (1) above has a synthetic resin portion and a first peptide portion. The peptide-containing resin (2) above has a synthetic resin portion and a second peptide portion. The peptide-containing resin (1) above preferably has a linker portion. In the peptide-containing resin (1) above, it is preferable that the synthetic resin portion and the first peptide portion are bound via the linker portion. The peptide-containing resin (2) above preferably has a linker portion. In the peptide-containing resin (2) above, it is preferable that the synthetic resin portion and the second peptide portion are bound via the linker portion. The peptide-containing resin (1) above may be used by one type only, or two or more types may be used in combination. The peptide-containing resin (2) described above may be used by one type only, or two or more types may be used in combination.

[0065] The preferred configuration of the synthetic resin portion in the peptide-containing resins (A), (1), and (2) described above will be explained below. Note that the common description of the synthetic resin portion in peptide-containing resin (A), the synthetic resin portion in peptide-containing resin (1), and the synthetic resin portion in peptide-containing resin (2) may simply be referred to as the "synthetic resin portion."

[0066] <Synthetic Resin Part> The above-mentioned synthetic resin part is a structural part derived from the synthetic resin in the peptide-containing resin (peptide-containing resin (A), (1), (2)). The type of the above-mentioned synthetic resin part is not particularly limited.

[0067] In the peptide-containing resin (A) described above, the synthetic resin portion is preferably a polyvinyl acetal resin portion or a (meth)acrylic polymer portion, and more preferably a polyvinyl acetal resin portion. In the peptide-containing resin (1) described above, the synthetic resin portion is preferably a polyvinyl acetal resin portion or a (meth)acrylic polymer portion, and more preferably a polyvinyl acetal resin portion. In the peptide-containing resin (2) described above, the synthetic resin portion is preferably a polyvinyl acetal resin portion or a (meth)acrylic polymer portion, and more preferably a polyvinyl acetal resin portion. In this case, memory T cells can be maintained more effectively, and T cells can be proliferated more effectively. Also, the dispersion component γ of the surface free energy ｄ and dipole component γ ｐ This makes it easier to adjust the range described above. Furthermore, it can improve the adhesion of cells to the cell scaffold material. The synthetic resin portion may also consist of both a polyvinyl acetal resin portion and a (meth)acrylic polymer portion.

[0068] In the peptide-containing resin (A) having the polyvinyl acetal resin portion described above, it is preferable that the polyvinyl acetal resin portion and the peptide portion are bonded via the linker portion. Therefore, it is preferable that the peptide-containing resin (A) having the polyvinyl acetal resin portion comprises a polyvinyl acetal resin portion, a linker portion, and a peptide portion.

[0069] In the peptide-containing resin (1) having the polyvinyl acetal resin portion described above, it is preferable that the polyvinyl acetal resin portion and the first peptide portion are bonded via the linker portion. Therefore, it is preferable that the peptide-containing resin (1) having the polyvinyl acetal resin portion has a polyvinyl acetal resin portion, a linker portion, and a first peptide portion.

[0070] In the peptide-containing resin (2) having the polyvinyl acetal resin portion described above, it is preferable that the polyvinyl acetal resin portion and the second peptide portion are bonded via the linker portion. Therefore, it is preferable that the peptide-containing resin (2) having the polyvinyl acetal resin portion has a polyvinyl acetal resin portion, a linker portion, and a second peptide portion.

[0071] In the peptide-containing resin (A) having the (meth)acrylic polymer portion described above, the (meth)acrylic polymer portion and the peptide portion may be bonded via a linker portion, or they may be directly bonded without a linker portion. The peptide-containing resin (A) having the (meth)acrylic polymer portion described above may also have a (meth)acrylic polymer portion, a linker portion, and a peptide portion.

[0072] In the peptide-containing resin (1) having the (meth)acrylic polymer portion described above, the (meth)acrylic polymer portion and the first peptide portion may be bonded via a linker portion, or they may be directly bonded without a linker portion. The peptide-containing resin (1) having the (meth)acrylic polymer portion may also have a (meth)acrylic polymer portion, a linker portion, and a first peptide portion.

[0073] In the peptide-containing resin (2) having the (meth)acrylic polymer portion described above, the (meth)acrylic polymer portion and the second peptide portion may be bonded via a linker portion, or they may be directly bonded without a linker portion. The peptide-containing resin (2) having the (meth)acrylic polymer portion may also have a (meth)acrylic polymer portion, a linker portion, and a second peptide portion.

[0074] Hereinafter, the polyvinyl acetal resin portion in peptide-containing resin (A), the polyvinyl acetal resin portion in peptide-containing resin (1), and the polyvinyl acetal resin portion in peptide-containing resin (2) will be described simply as "polyvinyl acetal resin portion." Similarly, the (meth)acrylic polymer portion in peptide-containing resin (A), the (meth)acrylic polymer portion in peptide-containing resin (1), and the (meth)acrylic polymer portion in peptide-containing resin (2) will be described simply as "(meth)acrylic polymer portion." In this specification, "(meth)acrylic" means either or both "acrylic" and "methacrylic," and "(meth)acrylate" means either or both "acrylate" and "methacrylate."

[0075] <<Polyvinyl acetal resin portion>> The above polyvinyl acetal resin portion (polyvinyl acetal resin skeleton) is a structural portion derived from polyvinyl acetal resin. The above synthetic resin portion is preferably a polyvinyl acetal resin portion. The above peptide-containing resin (A) preferably has a polyvinyl acetal resin portion and a peptide portion. The above peptide-containing resin (1) preferably has a polyvinyl acetal resin portion and a first peptide portion. The above peptide-containing resin (2) preferably has a polyvinyl acetal resin portion and a second peptide portion. The above cell scaffold material preferably contains a peptide-conjugated polyvinyl acetal resin having a polyvinyl acetal resin portion and a peptide portion. The above peptide-containing resin (A) preferably is a peptide-containing polyvinyl acetal resin. The above peptide-containing resin (1) preferably is a peptide-containing polyvinyl acetal resin. The above peptide-containing resin (2) preferably is a peptide-containing polyvinyl acetal resin. The above polyvinyl acetal resins may be used individually or in combination of two or more types.

[0076] The polyvinyl acetal resin portion described above preferably has structural units having acetal groups and structural units having hydroxyl groups, and more preferably has structural units having acetal groups, structural units having hydroxyl groups, and structural units having acetyl groups.

[0077] Polyvinyl acetal resin can be synthesized by acetalizing polyvinyl alcohol with an aldehyde.

[0078] The aldehyde used in the acetalization of polyvinyl alcohol is not particularly limited. Examples of the aldehyde include aldehydes having 1 to 10 carbon atoms. The aldehyde may or may not have a linear aliphatic group, a cyclic aliphatic group, or an aromatic group. The aldehyde may be a linear aldehyde or a cyclic aldehyde. Only one aldehyde may be used, or two or more may be used in combination.

[0079] The above-mentioned aldehyde is preferably formaldehyde, acetaldehyde, propionaldehyde, butyraldehyde, or pentanal, and more preferably butyraldehyde.

[0080] Therefore, the polyvinyl acetal resin portion is more preferably a polyvinyl butyral resin portion. The synthetic resin portion is more preferably a polyvinyl butyral resin portion. The peptide-containing resin (A) preferably has a polyvinyl butyral resin portion and a peptide portion. The peptide-containing resin (1) preferably has a polyvinyl butyral resin portion and a first peptide portion. The peptide-containing resin (2) preferably has a polyvinyl butyral resin portion and a second peptide portion. The cell scaffold material preferably contains a peptide-conjugated polyvinyl butyral resin having a polyvinyl butyral resin portion and a peptide portion. The peptide-containing resin (A) preferably is a peptide-containing polyvinyl butyral resin. The peptide-containing resin (1) preferably is a peptide-containing polyvinyl butyral resin. The peptide-containing resin (2) preferably is a peptide-containing polyvinyl butyral resin. In this case, memory T cells can be maintained more effectively, and T cells can be proliferated more effectively. Furthermore, the ability of cells to adhere to the cell scaffold material can be improved.

[0081] In the peptide-containing polyvinyl acetal resin (peptide-containing resin (A), (1), (2)) described above, the degree of acetalization A of the polyvinyl acetal resin portion (or the degree of butyralization in the case of the polyvinyl butyral resin portion) is preferably 40 mol% or more, more preferably 50 mol% or more, preferably 90 mol% or less, and more preferably 85 mol% or less. If the degree of acetalization A is above the lower limit, the peptide-containing polyvinyl acetal resin will not swell easily in the culture medium. If the degree of acetalization A is below the upper limit, good solubility in solvents can be achieved. Furthermore, if the degree of acetalization A is above the lower limit and below the upper limit, the dispersion component γ of the surface free energy ｄ and dipole component γ ｐ This makes it easier to adjust the range to the above-mentioned range.

[0082] In the peptide-containing polyvinyl acetal resins (peptide-containing resins (A), (1), (2)), the hydroxyl group content (hydroxyl group amount B) of the polyvinyl acetal resin portion is preferably 15 mol% or more, more preferably 20 mol% or more, preferably 45 mol% or less, more preferably 35 mol% or less, and even more preferably 30 mol% or less. When the hydroxyl group amount B is above the lower limit and below the upper limit, the dispersion component γ of the surface free energy ｄ and dipole component γ ｐ This makes it easier to adjust the range to the above-mentioned range.

[0083] In the above peptide-containing polyvinyl acetal resin (peptide-containing resin (A), (1), (2)), the degree of acetylation C (amount of acetyl groups) of the polyvinyl acetal resin portion is preferably 0.1 mol% or more, more preferably 0.5 mol% or more, preferably 5 mol% or less, and more preferably 4 mol% or less. When the degree of acetylation C is above the lower limit and below the upper limit, the reaction efficiency between the polyvinyl acetal resin and the linker can be increased. Furthermore, when the degree of acetylation C is above the lower limit and below the upper limit, the dispersion component γ of the surface free energy ｄ and dipole component γ ｐ This makes it easier to adjust the range to the above-mentioned range.

[0084] In the above-mentioned peptide-containing polyvinyl acetal resin, the degree of acetalization A of the polyvinyl acetal resin portion is usually the content (mol%) of structural units having acetal groups relative to the total content (100 mol%) of structural units constituting the peptide-containing polyvinyl acetal resin.

[0085] In the above-mentioned peptide-containing polyvinyl acetal resin, the amount of hydroxyl groups B in the polyvinyl acetal resin portion is usually the content (mol%) of structural units having hydroxyl groups relative to the total content (100 mol%) of structural units constituting the peptide-containing polyvinyl acetal resin.

[0086] In the above-mentioned peptide-containing polyvinyl acetal resin, the degree of acetylation C of the polyvinyl acetal resin portion is usually the content (mol%) of the structural units having the acetyl group relative to the total content (100 mol%) of the structural units constituting the peptide-containing polyvinyl acetal resin.

[0087] In the peptide-containing polyvinyl acetal resin described above, the content of structural units in the linker portion (for example, the content of structural units derived from polymerizable monomers) is defined as content L (mol%). In the peptide-containing polyvinyl acetal resin described above, the content of the peptide portion is defined as content P (mol%). The sum of the content of structural units constituting the peptide-containing polyvinyl acetal resin usually refers to the sum of the degree of acetalization A, the amount of hydroxyl groups B, the degree of acetylation C, content L, and content P.

[0088] The degree of acetalization A, the amount of hydroxyl groups B, and the degree of acetylation C are, １ It can be measured using 1H-NMR (nuclear magnetic resonance spectroscopy).

[0089] <<(meth)acrylic polymer portion>> The above (meth)acrylic polymer portion ((meth)acrylic polymer skeleton) is a structural portion derived from the (meth)acrylic polymer. The above synthetic resin portion is preferably the (meth)acrylic polymer portion. The above peptide-containing resin (A) preferably has a (meth)acrylic polymer portion and a peptide portion. The above peptide-containing resin (1) preferably has a (meth)acrylic polymer portion and a first peptide portion. The above peptide-containing resin (2) preferably has a (meth)acrylic polymer portion and a second peptide portion. The above cell scaffold material preferably contains a peptide-containing (meth)acrylic polymer having a (meth)acrylic polymer portion and a peptide portion. The above peptide-containing resin (A) is preferably a peptide-containing (meth)acrylic polymer. The above peptide-containing resin (1) is preferably a peptide-containing (meth)acrylic polymer. The above peptide-containing resin (2) is preferably a peptide-containing (meth)acrylic polymer. Only one type of (meth)acrylic polymer may be used, or two or more types may be used in combination.

[0090] The above-mentioned (meth)acrylic polymer part has a structural unit derived from a (meth)acrylate compound. Only one kind of the above-mentioned (meth)acrylate compound may be used, or two or more kinds may be used in combination.

[0091] The above-mentioned (meth)acrylic polymer part preferably has a structural unit derived from a (meth)acrylate compound (A) represented by the following formula (A). In this case, memory T cells can be maintained even better, and T cells can be proliferated even better. Also, the dispersive component γ of the above surface free energy ｄ and the dipole component γ ｐ can be more easily adjusted within the above range. Furthermore, the cell adhesion to the cell scaffold material can be enhanced. Only one kind of the (meth)acrylate compound (A) represented by the following formula (A) may be used, or two or more kinds may be used in combination.

[0092]

[0093] In the above formula (A), R １ represents a hydrogen atom or a methyl group, and R ２ represents a hydrocarbon group having 2 or more and 18 or less carbon atoms.

[0094] In the above formula (A), R １ may be a hydrogen atom or a methyl group.

[0095] R ２ in the above formula (A) may be an aliphatic hydrocarbon group or an aromatic hydrocarbon group. R ２ in the above formula (A) is preferably an aliphatic hydrocarbon group. The above aliphatic hydrocarbon group may be linear, may have a branched structure, may have a double bond, or may not have a double bond. R ２ in the above formula (A) may be an alkyl group or an alkylene group.

[0096] R ２The number of carbon atoms is preferably 4 or more, preferably 16 or less. When the number of carbon atoms is not less than the lower limit and not more than the upper limit, memory T cells can be maintained better, and T cells can proliferate better. Also, the dispersive component γ ｄ and the dipole component γ ｐ of the surface free energy are more easily adjusted within the above ranges. Furthermore, the cell adhesion to the cell scaffold material can be enhanced.

[0097] In 100 mol% of all the structural units of the (meth)acrylic polymer part, the content of the structural unit derived from the (meth)acrylate compound (A) is preferably 25 mol% or more, more preferably 30 mol% or more, preferably 98 mol% or less, more preferably 95 mol% or less. When the content is not less than the lower limit and not more than the upper limit, memory T cells can be maintained better, and T cells can proliferate better. Also, the dispersive component γ ｄ and the dipole component γ ｐ of the surface free energy are more easily adjusted within the above ranges. Furthermore, the cell adhesion to the cell scaffold material can be enhanced.

[0098] The (meth)acrylic polymer part preferably has a structural unit derived from a (meth)acrylate compound (B) different from the (meth)acrylate compound (A).

[0099] Examples of the (meth)acrylate compound (B) include (meth)acrylic acid, 3-butenoic acid, 4-pentenoic acid, 5-hexenoic acid, 6-heptenoic acid, 7-octenoic acid, benzenacrylic acid, (meth)acryloyloxyethyl succinic acid, (meth)acryloyloxyethyl phthalic acid, (meth)acryloyloxypropyl succinic acid, (meth)acryloyloxypropyl phthalic acid, (meth)acryloyloxyethyl hexahydro succinic acid, (meth)acryloyloxyethyl hexahydro phthalic acid, (meth)acryloyloxypropyl hexahydro succinic acid, and (meth)acryloyloxypropyl hexahydro phthalic acid.

[0100] The above (meth)acrylate compound (B) is preferably (meth)acrylic acid, (meth)acryloyloxyethyl succinic acid, (meth)acryloyloxypropyl succinic acid, (meth)acryloyloxyethyl hexahydrosuccinic acid, (meth)acryloyloxypropyl hexahydrosuccinic acid, or butenic acid, and more preferably (meth)acrylic acid. In this case, the effects of the present invention can be exhibited even more effectively.

[0101] Of the total structural units of the (meth)acrylic polymer portion described above, the content of structural units derived from the (meth)acrylate compound (B) is preferably 2 mol% or more, more preferably 5 mol% or more, more preferably 75 mol% or less, and more preferably 70 mol% or less. When the above content is above the lower limit and below the upper limit, memory T cells can be maintained more effectively, and T cells can be proliferated more effectively. Furthermore, the dispersion component γ of the surface free energy described above ｄ and dipole component γ ｐ This makes it easier to adjust the range described above. Furthermore, it can improve the adhesion of cells to the cell scaffold material.

[0102] In the total structural units of the (meth)acrylic polymer portion described above, the combined content of structural units derived from (meth)acrylate compound (A) and structural units derived from (meth)acrylate compound (B) is preferably 50 mol% or more, more preferably 65 mol% or more, even more preferably 80 mol% or more, still more preferably 90 mol% or more, particularly preferably 95 mol% or more, and most preferably 100 mol%. When the above combined content is above the above lower limit, memory T cells can be maintained more effectively, and T cells can be proliferated more effectively. Also, the dispersion component γ of the surface free energy ｄ and dipole component γ ｐ This makes it easier to adjust the concentration to the range described above. Furthermore, it can improve the adhesion of cells to the cell scaffold material. Note that the total concentration may be 100 mol% or less, or 90 mol% or less.

[0103] The preferred configurations of the peptide portions (first peptide portion, second peptide portion) in the peptide-containing resins (A), (1), and (2) described above will be explained below. Note that the first peptide portion in peptide-containing resin (A) and the first peptide portion in peptide-containing resin (1) may be simply referred to as the "first peptide portion." Similarly, the second peptide portion in peptide-containing resin (A) and the second peptide portion in peptide-containing resin (2) may be simply referred to as the "second peptide portion."

[0104] <Peptide portion> The peptide portions described above (first peptide portion, second peptide portion) are structural parts derived from peptides.

[0105] The peptide portion in the peptide-containing resin (A) has a first peptide portion and a second peptide portion. In the peptide-containing resin (A), the first peptide portion and the second peptide portion have different amino acid sequences. In the peptide-containing resin (A), the first peptide portion and the second peptide portion are not directly bonded. In the peptide-containing resin (A), one first peptide portion constitutes one structural unit. In the peptide-containing resin (A), one second peptide portion constitutes one structural unit. The first peptide portion and the second peptide portion have amino acid sequences. The peptide constituting the first peptide portion may be an oligopeptide or a polypeptide. The peptide constituting the second peptide portion may be an oligopeptide or a polypeptide. Only one type of peptide may be used, or two or more types may be used in combination.

[0106] The peptide-containing resin (1) has a first peptide portion. The peptide-containing resin (2) has a second peptide portion. The first peptide portion in the peptide-containing resin (1) and the second peptide portion in the peptide-containing resin (2) have different amino acid sequences. In the peptide-containing resin (1), one first peptide portion constitutes one structural unit. In the peptide-containing resin (2), one second peptide portion constitutes one structural unit. The first peptide portion and the second peptide portion have amino acid sequences. The peptide constituting the first peptide portion may be an oligopeptide or a polypeptide. The peptide constituting the second peptide portion may be an oligopeptide or a polypeptide. Only one type of peptide may be used, or two or more types may be used in combination.

[0107] The first peptide portion in the peptide-containing resin (A) is capable of binding to a first integrin, and the second peptide portion in the peptide-containing resin (A) is capable of binding to a second integrin. The first peptide portion in the peptide-containing resin (A) has an amino acid sequence capable of binding to a first integrin, and the second peptide portion in the peptide-containing resin (A) has an amino acid sequence capable of binding to a second integrin. It is preferable that the first integrin and the second integrin are different integrins.

[0108] The first peptide portion in the peptide-containing resin (1) is capable of binding to a first integrin, and the second peptide portion in the peptide-containing resin (2) is capable of binding to a second integrin. The first peptide portion in the peptide-containing resin (1) has an amino acid sequence capable of binding to a first integrin, and the second peptide portion in the peptide-containing resin (2) has an amino acid sequence capable of binding to a second integrin. It is preferable that the first integrin and the second integrin are different integrins.

[0109] Examples of integrins include integrin α5β1 (VLA-5), integrin αVβ1, integrin αVβ3, integrin αVβ5, integrin αVβ6, integrin αVβ8, integrin αIIbβ3, integrin α4β1 (VLA-4), integrin α4β7, integrin α9β1, integrin αDβ2, integrin αLβ2, integrin αMβ2, integrin αXβ2, and integrin αEβ7. For references on integrins, see Targeting integrin pathways: mechanisms and advances in therapy, Signal Transduction and Targeted Therapy volume 8, Article number: 1 (2023), URL: https: / / doi.org / 10.1038 / s41392-022-01259-6.

[0110] Preferably, the first integrin is integrin α5β1 (VLA-5) and the second integrin is integrin α4β1 (VLA-4). Preferably, the first peptide portion of the peptide-containing resin (A) is capable of binding to integrin α5β1 (VLA-5), and the second peptide portion of the peptide-containing resin (A) is capable of binding to integrin α4β1 (VLA-4). Preferably, the first peptide portion of the peptide-containing resin (1) is capable of binding to integrin α5β1 (VLA-5), and the second peptide portion of the peptide-containing resin (2) is capable of binding to integrin α4β1 (VLA-4). In this case, memory T cells can be maintained more effectively, and T cells can be proliferated more effectively.

[0111] The first peptide portion described above preferably has an RGD sequence (Arg-Gly-Asp). In this case, memory T cells can be maintained more effectively, and T cells can be proliferated more effectively. The peptide portion having the RGD sequence can be bound to VLA-5.

[0112] The second peptide portion described above preferably has an LDV sequence (Leu-Asp-Val). In this case, memory T cells can be maintained more effectively, and T cells can be proliferated more effectively. The peptide portion having the LDV sequence can be bound to VLA-4.

[0113] The first peptide portion described above may or may not have an LDV sequence, but it is preferable that it does not have an LDV sequence.

[0114] The second peptide portion described above may or may not have an RGD sequence, but it is preferable that it does not have an RGD sequence.

[0115] In the peptide-containing resin (A) described above, it is preferable that the synthetic resin portion and the first peptide portion are bonded via the linker portion, and that the synthetic resin portion and the second peptide portion are bonded via the linker portion. In addition, in the peptide-containing resin (A), there may be a structural portion in which the first peptide portion and the second peptide portion are bonded to the same linker portion.

[0116] The number of amino acid residues in the first peptide portion is preferably three or more, more preferably four or more, even more preferably five or more, preferably ten or fewer, more preferably eight or fewer, and even more preferably six or fewer. When the number of amino acid residues in the first peptide portion is above the lower limit and below the upper limit, memory T cells can be maintained more effectively, and T cells can be proliferated more effectively. Furthermore, the ability of cells to adhere to the cell scaffold material can be improved. The number of amino acid residues in the first peptide portion may exceed ten or exceed fifteen.

[0117] The number of amino acid residues in the second peptide portion is preferably three or more, more preferably four or more, even more preferably five or more, preferably ten or fewer, more preferably eight or fewer, and even more preferably six or fewer. When the number of amino acid residues in the second peptide portion is above the lower limit and below the upper limit, memory T cells can be maintained more effectively, and T cells can be proliferated more effectively. Furthermore, the ability of cells to adhere to the cell scaffold material can be improved. The number of amino acid residues in the second peptide portion may exceed ten or exceed fifteen.

[0118] The first peptide portion described above may be linear or may have a cyclic peptide skeleton. The cyclic peptide skeleton is a cyclic skeleton composed of multiple amino acids. It is preferable that the first peptide portion has a cyclic peptide skeleton. In this case, memory T cells can be maintained more effectively, and T cells can be proliferated more effectively. Furthermore, the ability of cells to adhere to the cell scaffold material can be improved.

[0119] When the first peptide portion has the cyclic peptide skeleton, the cyclic peptide skeleton is preferably composed of four or more amino acids, more preferably of five or more amino acids, and preferably of ten or fewer amino acids.

[0120] The second peptide portion may be linear or may have a cyclic peptide skeleton. It is preferable that the second peptide portion is linear. It is preferable that the second peptide portion does not have a cyclic peptide skeleton. In this case, memory T cells can be maintained more effectively, and T cells can be proliferated more effectively. Furthermore, the ability of cells to adhere to the cell scaffold material can be improved.

[0121] The first peptide portion described above preferably has an RGD sequence represented by the following formula (P1), and more preferably has a c-RGDfK sequence. In this case, memory T cells can be maintained more effectively, and T cells can be proliferated more effectively. Furthermore, the ability of cells to adhere to the cell scaffold material can be improved. The c-RGDfK sequence is an amino acid sequence of Arg-Gly-Asp-Phe-Lys, in which Arg and Lys are bound together to form a cyclic peptide skeleton, and Phe is the D-isomer.

[0122] Arg-Gly-Asp-X...Formula (P1)

[0123] In the above formula (P1), X represents Gly, Ala, Val, Ser, Thr, Phe, Met, Pro, or Asn.

[0124] The second peptide portion described above preferably has an EILDV sequence (Glu-Ile-Leu-Asp-Val). In this case, memory T cells can be maintained more effectively, and T cells can be proliferated more effectively. Furthermore, the ability of cells to adhere to the cell scaffold material can be improved.

[0125] In the peptide-containing resin (A) described above, the content P1 of the first peptide portion is preferably 0.01 mol% or more, more preferably 0.02 mol% or more, even more preferably 0.03 mol% or more, particularly preferably 0.04 mol% or more, preferably 2.0 mol% or less, more preferably 1.7 mol% or less, even more preferably 1.5 mol% or less, and particularly preferably 1.2 mol% or less. When the content P1 is above the lower limit and below the upper limit, memory T cells can be maintained more effectively, and T cells can be proliferated more effectively. Furthermore, the ability of cells to adhere to the cell scaffold material can be improved.

[0126] In the peptide-containing resin (1) described above, the content P1 of the first peptide portion is preferably 0.01 mol% or more, more preferably 0.02 mol% or more, even more preferably 0.03 mol% or more, particularly preferably 0.04 mol% or more, preferably 2.0 mol% or less, more preferably 1.7 mol% or less, even more preferably 1.3 mol% or less, and particularly preferably 1.0 mol% or less. When the content P1 is above the lower limit and below the upper limit, memory T cells can be maintained more effectively, and T cells can be proliferated more effectively. Furthermore, the ability of cells to adhere to the cell scaffold material can be improved.

[0127] In the peptide-containing resin (A) described above, the content P2 of the second peptide portion is preferably 0.01 mol% or more, more preferably 0.03 mol% or more, even more preferably 0.04 mol% or more, particularly preferably 0.06 mol% or more, preferably 2.0 mol% or less, more preferably 1.8 mol% or less, even more preferably 1.5 mol% or less, and particularly preferably 1.2 mol% or less. When the content P2 is above the lower limit and below the upper limit, memory T cells can be maintained more effectively, and T cells can be proliferated more effectively. Furthermore, the ability of cells to adhere to the cell scaffold material can be improved.

[0128] In the peptide-containing resin (2) described above, the content P2 of the second peptide portion is preferably 0.01 mol% or more, more preferably 0.03 mol% or more, even more preferably 0.04 mol% or more, particularly preferably 0.06 mol% or more, preferably 2.0 mol% or less, more preferably 1.8 mol% or less, even more preferably 1.5 mol% or less, and particularly preferably 1.2 mol% or less. When the content P2 is above the lower limit and below the upper limit, memory T cells can be maintained more effectively, and T cells can be proliferated more effectively. Furthermore, the ability of cells to adhere to the cell scaffold material can be improved.

[0129] In the peptide-containing resin (A) described above, the sum of the content P1 of the first peptide portion and the content P2 of the second peptide portion is preferably 0.02 mol% or more, more preferably 0.05 mol% or more, even more preferably 0.07 mol% or more, particularly preferably 0.1 mol% or more, preferably 4.0 mol% or less, more preferably 3.5 mol% or less, even more preferably 3.3 mol% or less, and particularly preferably 3.0 mol% or less. When the sum is above the lower limit, memory T cells can be maintained more effectively, and T cells can be proliferated more effectively. Furthermore, the ability of cells to adhere to the cell scaffold material can be improved. When the sum is below the upper limit, solubility in solvents can be improved.

[0130] The sum of the content P1 of the first peptide portion in the peptide-containing resin (1) and the content P2 of the second peptide portion in the peptide-containing resin (2) is preferably 0.02 mol% or more, more preferably 0.05 mol% or more, even more preferably 0.07 mol% or more, particularly preferably 0.1 mol% or more, preferably 4.0 mol% or less, more preferably 3.5 mol% or less, even more preferably 3.3 mol% or less, and particularly preferably 3.0 mol% or less. When the sum is above the lower limit, memory T cells can be maintained more effectively, and T cells can be proliferated more effectively. Furthermore, the ability of cells to adhere to the cell scaffold material can be improved. When the sum is below the upper limit, solubility in the solvent can be improved.

[0131] In the peptide-containing resin (A) described above, the molar ratio (content P1 / content P2) of the content P1 of the first peptide portion to the content P2 of the second peptide portion is preferably 0.03 or more, more preferably 0.07 or more, even more preferably 0.14 or more, preferably 25 or less, even more preferably 15 or less, and even more preferably 7 or less. When the molar ratio (content P1 / content P2) is above the lower limit and below the upper limit, memory T cells can be maintained more effectively, and T cells can be proliferated more effectively. Furthermore, the ability of cells to adhere to the cell scaffold material can be improved.

[0132] The molar ratio (content P1 / content P2) of the content P1 of the first peptide portion in the peptide-containing resin (1) to the content P2 of the second peptide portion in the peptide-containing resin (2) is preferably 0.03 or more, more preferably 0.07 or more, even more preferably 0.14 or more, preferably 25 or less, even more preferably 15 or less, and even more preferably 7 or less. When the molar ratio (content P1 / content P2) is above the lower limit and below the upper limit, the efficiency of molecular introduction into cells can be further increased. In addition, the ability of cells to adhere to the cell scaffold material can be improved, and the efficiency of cell proliferation can be increased.

[0133] In the peptide-containing resin (A) described above, the molar ratio ((content P1 + content P2) / content P) of the sum of the content P1 of the first peptide portion and the content P2 of the second peptide portion to the content P of the peptide portion is preferably 0.5 or more, more preferably 0.6 or more, even more preferably 0.7 or more, still more preferably 0.8 or more, particularly preferably 0.9 or more, and most preferably 1. When the molar ratio ((content P1 + content P2) / content P) is above the lower limit, memory T cells can be maintained more effectively, and T cells can be proliferated more effectively. Furthermore, the ability of cells to adhere to the cell scaffold material can be improved. In addition, in the peptide-containing resin (A) described above, the molar ratio ((content P1 + content P2) / content P) may be 1 or less, or less than 1.

[0134] The above content P is the content (mol%) of the peptide portion relative to the total content (100 mol%) of the structural units constituting the peptide-containing resin. The above content P1 is the content (mol%) of the first peptide portion relative to the total content (100 mol%) of the structural units constituting the peptide-containing resin. The above content P2 is the content (mol%) of the second peptide portion relative to the total content (100 mol%) of the structural units constituting the peptide-containing resin.

[0135] The content P of the peptide portion, the content P1 of the first peptide portion, and the content P2 of the second peptide portion can be measured, for example, by NMR (nuclear magnetic resonance).

[0136] In the case where the peptide-containing resin (A) described above is a peptide-containing resin (peptide-containing polyvinyl acetal resin) having a polyvinyl acetal resin portion and a linker portion, a preferred configuration of the peptide portion content will be further explained.

[0137] As described above, in the peptide-containing polyvinyl acetal resin, the degree of acetalization of the polyvinyl acetal resin portion is denoted as degree of acetalization A, the hydroxyl group content of the polyvinyl acetal resin portion is denoted as hydroxyl group content B, and the degree of acetylation of the polyvinyl acetal resin portion is denoted as degree of acetylation C. Also, as described above, in the peptide-containing polyvinyl acetal resin, the content of structural units in the linker portion (for example, the content of structural units derived from polymerizable monomers) is denoted as content L. Also, as described above, in the peptide-containing polyvinyl acetal resin, the content of the first peptide portion is denoted as content P1, and the content of the second peptide portion is denoted as content P2.

[0138] In the peptide-containing polyvinyl acetal resin (peptide-containing resin (A)) described above, of the total of acetalization degree A, hydroxyl group amount B, acetylation degree C, content L, content P1, and content P2 in 100 mol%, content P1 is preferably 0.01 mol% or more, more preferably 0.02 mol% or more, even more preferably 0.03 mol% or more, particularly preferably 0.04 mol% or more, preferably 2.0 mol% or less, more preferably 1.7 mol% or less, even more preferably 1.5 mol% or less, and particularly preferably 1.2 mol% or less. When the above content P1 is above the lower limit and below the upper limit, memory T cells can be maintained more effectively, and T cells can be proliferated more effectively. Furthermore, the ability of cells to adhere to the cell scaffold material can be improved.

[0139] In the peptide-containing polyvinyl acetal resin (peptide-containing resin (A)) described above, of the total of acetalization degree A, hydroxyl group amount B, acetylation degree C, content L, content P1, and content P2 in 100 mol%, content P2 is preferably 0.01 mol% or more, more preferably 0.03 mol% or more, even more preferably 0.05 mol% or more, particularly preferably 0.07 mol% or more, preferably 2.0 mol% or less, more preferably 1.8 mol% or less, even more preferably 1.6 mol% or less, and particularly preferably 1.4 mol% or less. When the above content P2 is above the lower limit and below the upper limit, memory T cells can be maintained more effectively, and T cells can be proliferated more effectively. Furthermore, the ability of cells to adhere to the cell scaffold material can be improved.

[0140] In the peptide-containing polyvinyl acetal resin (peptide-containing resin (A)) described above, the sum of the acetalization degree A, hydroxyl group amount B, acetylation degree C, content L, content P1, and content P2 in a total of 100 mol%, is preferably 0.02 mol% or more, more preferably 0.05 mol% or more, even more preferably 0.07 mol% or more, particularly preferably 0.10 mol% or more, preferably 4.0 mol% or less, more preferably 3.5 mol% or less, even more preferably 3.0 mol% or less, and particularly preferably 2.5 mol% or less. When the above sum is above the lower limit and below the upper limit, memory T cells can be maintained more effectively, and T cells can be proliferated more effectively. Furthermore, the ability of cells to adhere to the cell scaffold material can be improved.

[0141] The preferred configuration of the linker portion in the peptide-containing resins (A), (1), and (2) described above will be explained below. Note that the linker portion in peptide-containing resin (A), the linker portion in peptide-containing resin (1), and the linker portion in peptide-containing resin (2) will sometimes be simply referred to as the "linker portion."

[0142] <Linker portion> The linker portion is a structural portion derived from the linker. The linker portion is formed by the linker. The peptide-containing resin (A) preferably has a linker portion. The peptide-containing resin (1) preferably has a linker portion. The peptide-containing resin (2) preferably has a linker portion. Only one type of linker may be used, or two or more types may be used in combination.

[0143] The linker is preferably a polymerizable monomer. Therefore, the linker portion in the peptide-containing resin (A) preferably has structural units derived from a polymerizable monomer. The linker portion in the peptide-containing resin (1) preferably has structural units derived from a polymerizable monomer. The linker portion in the peptide-containing resin (2) preferably has structural units derived from a polymerizable monomer. Only one polymerizable monomer may be used, or two or more may be used in combination.

[0144] Examples of polymerizable monomers include (meth)acrylate compounds. Examples of (meth)acrylate compounds include the (meth)acrylate compound (A) and the (meth)acrylate compound (B) described above.

[0145] From the viewpoint of good reaction with peptides, the polymerizable monomer is preferably a (meth)acrylate compound. The linker portion in the peptide-containing resin (A) preferably has structural units derived from a (meth)acrylate compound. The linker portion in the peptide-containing resin (1) preferably has structural units derived from a (meth)acrylate compound. The linker portion in the peptide-containing resin (2) preferably has structural units derived from a (meth)acrylate compound.

[0146] The polymerizable monomer described above may or may not have an aromatic skeleton.

[0147] The polymerizable monomer is preferably a compound having a functional group that can bind to the peptide, and more preferably a compound having a functional group that can condense with the carboxyl group or amino group of the peptide.

[0148] Functional groups that can condense with the carboxyl group or amino group of the above peptide include carboxyl groups, thiol groups, amino groups, hydroxyl groups, and cyano groups.

[0149] From the viewpoint of reacting well with peptides, the polymerizable monomer preferably contains a compound having a carboxyl group or an amino group, and more preferably contains a compound having a carboxyl group. The linker portion in the peptide-containing resin (A) preferably has structural units derived from a compound having a carboxyl group. The linker portion in the peptide-containing resin (1) preferably has structural units derived from a compound having a carboxyl group. The linker portion in the peptide-containing resin (2) preferably has structural units derived from a compound having a carboxyl group.

[0150] When obtaining a peptide-containing resin having a polyvinyl acetal resin portion, examples of the carboxyl group-containing compound (carboxylic acid monomer) include (meth)acrylic acid and carboxyl group-containing acrylamide. By using the above carboxyl group-containing compound (carboxylic acid monomer), the carboxylic acid monomer can be polymerized by graft polymerization when a linker is introduced, thereby increasing the number of carboxyl groups that can react with the peptide.

[0151] In the peptide-containing resin (A) described above, the linker portion preferably has structural units derived from a (meth)acrylate compound that does not have a carboxyl group and structural units derived from a (meth)acrylate compound that has a carboxyl group. In the peptide-containing resin (1) described above, the linker portion preferably has structural units derived from a (meth)acrylate compound that does not have a carboxyl group and structural units derived from a (meth)acrylate compound that has a carboxyl group. In the peptide-containing resin (2) described above, the linker portion preferably has structural units derived from a (meth)acrylate compound that does not have a carboxyl group and structural units derived from a (meth)acrylate compound that has a carboxyl group. In this case, the dispersion component γ of the surface free energy ｄ and dipole component γ ｐ This makes it easier to adjust the range to the above-mentioned range.

[0152] In the peptide-containing resin (A) described above, it is preferable that the structural units derived from the (meth)acrylate compound that does not have a carboxyl group include structural units derived from butyl (meth)acrylate. In the peptide-containing resin (1) described above, it is preferable that the structural units derived from the (meth)acrylate compound that does not have a carboxyl group include structural units derived from butyl (meth)acrylate. In the peptide-containing resin (2) described above, it is preferable that the structural units derived from the (meth)acrylate compound that does not have a carboxyl group include structural units derived from butyl (meth)acrylate. In this case, the dispersion component γ of the surface free energy ｄ and dipole component γ ｐ This makes it easier to adjust the range to the above-mentioned range.

[0153] In the peptide-containing resin (A) described above, the structural units derived from the (meth)acrylate compound having a carboxyl group are more preferably derived from (meth)acrylic acid, and even more preferably from acrylic acid. In the peptide-containing resin (1) described above, the structural units derived from the (meth)acrylate compound having a carboxyl group are more preferably derived from (meth)acrylic acid, and even more preferably from acrylic acid. In the peptide-containing resin (2) described above, the structural units derived from the (meth)acrylate compound having a carboxyl group are more preferably derived from (meth)acrylic acid, and even more preferably from acrylic acid. In this case, the synthetic resin portion and the peptide portion can be well bonded via the linker portion.

[0154] When obtaining a peptide-containing resin having a (meth)acrylic polymer portion, it is preferable that the polymerizable monomer has a functional group capable of binding to a (meth)acrylic acid ester. Examples of functional groups capable of binding to a (meth)acrylic acid ester include vinyl groups, (meth)acryloyl groups, and allyl groups. It is preferable that the polymerizable monomer has a (meth)acryloyl group, and more preferably has a carboxyl group or an amino group, and also has a (meth)acryloyl group.

[0155] Examples of polymerizable monomers used to obtain a peptide-containing resin having a (meth)acrylic polymer portion include (meth)acrylic acid, itaconic acid, and acrylamide.

[0156] From the viewpoint of effectively bonding the (meth)acrylic polymer and the peptide via the linker portion, the polymerizable monomer is preferably (meth)acrylic acid or itaconic acid, and more preferably (meth)acrylic acid.

[0157] <Further Details of Peptide-Containing Resins> The number average molecular weight of the peptide-containing resins (peptide-containing resins (A), (1), (2)) is preferably 10,000 or more, more preferably 50,000 or more, even more preferably 100,000 or more, preferably 5,000,000 or less, more preferably 2,500,000 or less, and even more preferably 1,000,000 or less. If the number average molecular weight is above the lower limit, the cell scaffold material will be less likely to dissolve into the liquid culture medium during cell culture, and the cell scaffold material will be less likely to peel off the main body of the substrate during cell culture. If the number average molecular weight is below the upper limit, the solubility in alcohol solvents can be increased, making it easier to coat the main body of the substrate with a composition containing the cell scaffold material and alcohol solvent.

[0158] The number-average molecular weight of the peptide-containing resin can be measured, for example, by the following method. The peptide-containing resin is dissolved in tetrahydrofuran (THF) to prepare a 0.2% by weight solution of the peptide-containing resin. Next, it is evaluated using a gel permeation chromatography (GPC) analyzer (APC system, Waters Corporation) under the following measurement conditions.

[0159] Column: HSPgel HR MB-M 6.0 × 150 mm Flow rate: 0.5 mL / min Column temperature: 40°C Injection volume: 10 μL Detector: RI, PDA Standard sample: Polystyrene

[0160] Examples of methods for obtaining a peptide-containing resin (peptide-containing polyvinyl acetal resin) having the above-mentioned polyvinyl acetal resin portion include the following methods.

[0161] A polyvinyl acetal resin is reacted with a linker (polymerizable monomer) to obtain a reaction product in which the polyvinyl acetal resin and the linker are bonded. The obtained reaction product is then reacted with a peptide to obtain a peptide-containing polyvinyl acetal resin.

[0162] Methods for obtaining a peptide-containing resin (peptide-containing (meth)acrylic polymer) having the above-mentioned (meth)acrylic polymer portion include, for example, the following methods (1) and (2).

[0163] Method (1): A (meth)acrylic polymer is reacted with a linker (polymerizable monomer) to obtain a reaction product in which the (meth)acrylic polymer and the linker are bonded. The obtained reaction product is reacted with a peptide to obtain a peptide-containing (meth)acrylic polymer.

[0164] Method (2): A peptide-containing (meth)acrylic polymer is obtained by reacting a (meth)acrylic polymer with a peptide.

[0165] From the viewpoint of maintaining memory T cells more effectively and promoting T cell proliferation more effectively, it is preferable that the peptide-containing resin (A) has a main chain and a graft chain. In this case, it is preferable that the synthetic resin portion constitutes the main chain. It is also preferable that the graft chain has a structural unit derived from the polymerizable monomer and the peptide portion. It is preferable that the graft chain has a linker portion bound to the main chain and a peptide portion bound to the linker portion. It is preferable that the graft chain has a linker portion bound to the main chain, a first peptide portion bound to the linker portion and a second peptide portion bound to the linker portion.

[0166] From the viewpoint of maintaining memory T cells more effectively and promoting T cell proliferation more effectively, it is preferable that the peptide-containing resin (1) has a main chain and a graft chain. In this case, it is preferable that the synthetic resin portion constitutes the main chain. It is also preferable that the graft chain has a structural unit derived from the polymerizable monomer and the first peptide portion. It is preferable that the graft chain has a linker portion bound to the main chain and the first peptide portion bound to the linker portion.

[0167] From the viewpoint of maintaining memory T cells more effectively and promoting T cell proliferation more effectively, it is preferable that the peptide-containing resin (2) has a main chain and a graft chain. In this case, it is preferable that the synthetic resin portion constitutes the main chain. It is also preferable that the graft chain has a structural unit derived from the polymerizable monomer and the second peptide portion. It is preferable that the graft chain has a linker portion bound to the main chain and the second peptide portion bound to the linker portion.

[0168] Figure 1 is a schematic diagram showing an example of a peptide-containing resin (A) that can be used in the present invention.

[0169] The peptide-containing resin 10 shown in Figure 1 is the peptide-containing resin (A) described above. The peptide-containing resin 10 has a synthetic resin portion 11, a linker portion 12, and a peptide portion 15. The peptide portion 15 has a first peptide portion 13 and a second peptide portion 14. Therefore, the peptide-containing resin 10 has a synthetic resin portion 11, a linker portion 12, a first peptide portion 13, and a second peptide portion 14. In Figure 1, to facilitate understanding the similarities and differences between the linker portion 12, the first peptide portion 13, and the second peptide portion 14, the first peptide portion 13 is represented by a circle and the second peptide portion 14 is represented by a triangle.

[0170] The first peptide portion 13 has an RGD sequence. The second peptide portion 14 has an LDV sequence. The peptide-containing resin 10 has a main chain and a graft chain. The synthetic resin portion 11 constitutes the main chain of the peptide-containing resin 10. The graft chain has a linker portion 12, a first peptide portion 13, and a second peptide portion 14. The linker portion 12 is bound to the synthetic resin portion 11. The first peptide portion 13 is bound to the linker portion 12. The second peptide portion 14 is bound to the linker portion 12. The synthetic resin portion 11 and the first peptide portion 13 are bound via the linker portion 12, and the synthetic resin portion 11 and the second peptide portion 14 are bound via the linker portion 12. In the peptide-containing resin 10, the first peptide portion 13 and the second peptide portion 14 are bound to the same linker portion 12.

[0171] Figure 2(a) is a schematic diagram showing an example of a peptide-containing resin (1) that can be used in the present invention, and Figure 2(b) is a schematic diagram showing an example of a peptide-containing resin (2) that can be used in the present invention.

[0172] The peptide-containing resin 10a shown in Figure 2(a) is the peptide-containing resin (1) described above. The peptide-containing resin 10a has a synthetic resin portion 11a, a linker portion 12a, and a first peptide portion 13a. In Figure 2(a), the first peptide portion 13a is shown as a circle to facilitate understanding the similarities and differences between the linker portion 12a and the first peptide portion 13a.

[0173] The first peptide portion 13a has an RGD sequence. The peptide-containing resin 10a has a main chain and a graft chain. The synthetic resin portion 11a constitutes the main chain of the peptide-containing resin 10a. The graft chain has a linker portion 12a and the first peptide portion 13a. The linker portion 12a is bound to the synthetic resin portion 11a. The first peptide portion 13a is bound to the linker portion 12a. The synthetic resin portion 11a and the first peptide portion 13a are linked via the linker portion 12a.

[0174] The peptide-containing resin 10b shown in Figure 2(b) is the peptide-containing resin (2) described above. The peptide-containing resin 10b has a synthetic resin portion 11b, a linker portion 12b, and a second peptide portion 14b. In Figure 2(b), the second peptide portion 14b is represented by a triangle to facilitate understanding the similarities and differences between the linker portion 12b and the second peptide portion 14b.

[0175] The second peptide portion 14b has an LDV sequence. The peptide-containing resin 10b has a main chain and a graft chain. The synthetic resin portion 11b constitutes the main chain of the peptide-containing resin 10b. The graft chain has a linker portion 12b and the second peptide portion 14b. The linker portion 12b is bound to the synthetic resin portion 11b. The second peptide portion 14b is bound to the linker portion 12b. The synthetic resin portion 11b and the second peptide portion 14b are linked via the linker portion 12b.

[0176] (Further details of cell scaffold material) The cell scaffold material (A) above comprises the peptide-containing resin (A). The cell scaffold material (B) above comprises the peptide-containing resin (1) and the peptide-containing resin (2). The cell scaffold material may or may not contain other components other than the peptide-containing resin. The cell scaffold material may or may not contain, for example, polyvinyl acetal resin (polyvinyl acetal resin without peptides). The cell scaffold material may or may not contain, for example, (meth)acrylic polymer ((meth)acrylic polymer without peptides). Other components may include polyolefin resin, polyether resin, polyester, epoxy resin, polyamide resin, polyimide resin, polyurethane resin, polycarbonate resin, cellulose, and polypeptide. Only one of the other components may be used, or two or more may be used in combination.

[0177] From the viewpoint of improving the adhesion of cells to the cell scaffold material, cell scaffold material (A) containing peptide-containing resin (A) is more preferable to cell scaffold material (B) containing peptide-containing resins (1) and (2). This is because cell scaffold material (A) contains peptide-containing resin (A) which has a first and second peptide portion in one molecule, and therefore these first and second peptide portions are likely to be adjacent to each other.

[0178] In 100% by weight of the cell scaffold material (A), the content of the peptide-containing resin (A) is preferably 90% by weight or more, more preferably 95% by weight or more, even more preferably 97.5% by weight or more, particularly preferably 99% by weight or more, and most preferably 100% by weight (total amount). When the content of the peptide-containing resin (A) is above the lower limit, the effects of the present invention can be exhibited even more effectively. In 100% by weight of the cell scaffold material (A), the content of the peptide-containing resin (A) may be 100% by weight or less, or less than 100% by weight.

[0179] In 100% by weight of the cell scaffold material (B), the total content of the peptide-containing resin (1) and the peptide-containing resin (2) is preferably 90% by weight or more, more preferably 95% by weight or more, even more preferably 97.5% by weight or more, particularly preferably 99% by weight or more, and most preferably 100% by weight (total amount). When the total content is above the lower limit, the effects of the present invention can be exhibited even more effectively. In 100% by weight of the cell scaffold material (B), the total content of the peptide-containing resin (1) and the peptide-containing resin (2) may be 100% by weight or less, or less than 100% by weight.

[0180] The above cell scaffold material is used to culture a cell population containing T cells. The above cell scaffold material is used at least to culture a cell population containing T cells.

[0181] Examples of the above-mentioned T cells include helper T cells, regulatory T cells, killer T cells, natural killer T cells, and γΔT cells. Only one type of T cell may be used, or two or more types may be used in combination. Furthermore, the differentiation phase of the above-mentioned T cells is not particularly limited. The above-mentioned T cells (or the differentiation phase of the above-mentioned T cells) may be naive T cells, memory T cells, effector T cells, or exhausted T cells. The above-mentioned T cells may be T cells with only one differentiation phase, or T cells with different differentiation phases may be used in combination.

[0182] The above-mentioned T cells preferably include memory T cells. Examples of memory T cells include stem cell memory T cells, central memory T cells, and effector memory T cells.

[0183] The above T cells preferably include CD8+ T cells. In this case, memory T cells can be maintained more effectively, and T cells can be proliferated more effectively.

[0184] The T cells mentioned above may be donor-derived T cells, T cells obtained by culturing donor-derived T cells, or iPS cell-derived T cells. The T cells mentioned above may also be gene-transformed T cells. Furthermore, the T cells mentioned above may be cytokine-stimulated T cells (cytokine-activated T cells).

[0185] [Cell Culture Substrate] The cell culture substrate according to the present invention comprises a substrate body and a cell scaffold material disposed on the first surface of the substrate body. In the cell culture substrate according to the present invention, the cell scaffold material is the cell scaffold material described above.

[0186] The material, size, and shape of the substrate body described above are not particularly limited. Conventional known substrate bodies can be used as the substrate body.

[0187] Examples of materials for the main body of the substrate include synthetic resins, metals, and glass. Examples of synthetic resins include polystyrene, polyethylene, polyethylene terephthalate, polypropylene, polyethersulfone, polycarbonate, polyester, polyisoprene, cycloolefin polymer, polyimide, polyamide, polyamideimide, (meth)acrylic resin, epoxy resin, and silicone.

[0188] The substrate body can take the form of fibrous material, particulate material, plate-like material, film-like material, porous membrane-like material, or bag-like material. The substrate body may also be a container.

[0189] The above-mentioned substrate body is preferably a container. The shape and size of the above-mentioned container (substrate body) are not particularly limited. Examples of the above-mentioned container (substrate body) include 2 to 384 well plates, single-walled flasks, multi-walled flasks, multifaceted flasks, dishes, roller bottles, bags, insert cups, and microfluidic tips.

[0190] Figure 3 is a schematic cross-sectional view showing a cell culture substrate according to one embodiment of the present invention.

[0191] The cell culture substrate 1 comprises a substrate body 2 and a cell scaffold material 3. The substrate body 2 is a container. The cell culture substrate 1 is a cell culture vessel. The cell scaffold material 3 is arranged on the first surface 2a of the substrate body 2.

[0192] [Method for culturing T cells] The method for culturing T cells according to the present invention comprises a culture step of culturing a cell population including T cells on the surface of the cell scaffold material described above.

[0193] The above-described method for culturing T cells is a method using the cell culture substrate described above, and preferably comprises a culture step of culturing a cell population including T cells on the surface of the cell scaffold material in the cell culture substrate.

[0194] The above culture step is a step of culturing a cell population containing T cells on the surface of the cell scaffold material. The above culture step is a step of adhering culture of the cell population containing T cells. In the above culture step, a cell population containing T cells can be cultured under conventionally known conditions, except that the cell scaffold material according to the present invention is used as the cell scaffold material.

[0195] The above method for culturing T cells preferably includes a step of bringing a cell population containing T cells into contact with cytokines. The above method for culturing T cells preferably includes a first culture step of culturing a cell population containing T cells on the surface of the cell scaffold material, a step of bringing a cell population containing T cells attached to the surface of the cell scaffold material into contact with cytokines, and a second culture step of culturing a cell population containing T cells stimulated by the cytokines on the surface of the cell scaffold material.

[0196] Examples of the cytokines mentioned above include interleukin-2 (IL-2), IL-7, and IL-15. These cytokines may be used individually or in combination of two or more.

[0197] The present invention will be specifically described below with reference to examples and comparative examples. The present invention is not limited to the following examples. The content of structural units in the obtained peptide-containing resin, for example, the degree of acetalization A, the amount of hydroxyl groups B, the degree of acetylation C, the content of structural units derived from polymerizable monomers, the content of the first peptide portion P1, and the content of the second peptide portion P2 are determined by dissolving the peptide-containing resin in DMSO-d6 (dimethyl sulfoxide). １ The measurements were performed using 1H-NMR (nuclear magnetic resonance spectroscopy).

[0198] (Example 1) (Preparation of Polyvinyl Acetal Resin X) 2700 mL of deionized water and 300 g of polyvinyl alcohol with an average degree of polymerization of 250 and a degree of saponification of 99 mol% were added to a reactor equipped with a stirring device, and the mixture was heated and dissolved while stirring to obtain a solution. Next, 35 wt% hydrochloric acid was added to this solution as a catalyst to a hydrochloric acid concentration of 0.2 wt%, and the temperature was adjusted to 15°C. Then, 22 g of n-butyraldehyde (n-BA) was added while stirring. Subsequently, 148 g of n-butyraldehyde (n-BA) was added, and white particulate polyvinyl acetal resin precipitated. Fifteen minutes after precipitation, 35 wt% hydrochloric acid was added to a hydrochloric acid concentration of 1.8 wt%, the mixture was heated to 50°C, and aged at 50°C for 2 hours. Next, the solution was cooled and neutralized, then the polyvinyl acetal resin was washed with water and dried to obtain polyvinyl acetal resin (polyvinyl butyral resin). A mixed solution was obtained by dissolving 6 parts by weight of acrylic acid, 21 parts by weight of butyl acrylate, and 70 parts by weight of the obtained polyvinyl acetal resin in 255 parts by weight of tetrahydrofuran. 0.015 parts by weight of perbutyl O (manufactured by NOF Corporation) was dissolved in the obtained mixed solution and reacted at 90°C for 6 hours. Next, the reaction solution was mixed with 30,000 parts by weight of water. The obtained precipitate was vacuum dried at 80°C for 3 hours to produce polyvinyl acetal resin X (polyvinyl acetal resin with a linker portion) having structural units derived from acrylic acid and structural units derived from butyl acrylate.

[0199] (Preparation Steps) DMF was prepared as the first solvent. DMF was prepared as the second solvent. A cyclic peptide having the amino acid sequence Arg-Gly-Asp-Phe-Lys (5 amino acid residues, Arg and Lys combine to form a cyclic skeleton, Phe is the D-isomer, c-RGDfK) was prepared as the first peptide. A peptide having the amino acid sequence Glu-Ile-Leu-Asp-Val (5 amino acid residues, EILDV) was prepared as the second peptide. 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride was prepared as the condensing agent. The first solution was prepared by mixing 50 parts by weight of polyvinyl acetal resin X, 3 parts by weight of the first peptide, and 3 parts by weight of the second peptide with 1000 parts by weight of the first solvent. The second solution was prepared by mixing 10 parts by weight of the condensing agent with 1000 parts by weight of the second solvent. The first solution and the second solution were mixed to prepare a solution containing polyvinyl acetal resin X, the first peptide, the second peptide, and a condensing agent.

[0200] (Reaction step) The obtained solution was reacted at 40°C for 2 hours to dehydrate and condense the carboxyl groups in the structural units derived from acrylic acid of polyvinyl acetal resin X with the amino groups of Lys of the first peptide or Glu of the second peptide, thereby obtaining a solution containing peptide-containing polyvinyl acetal resin (A).

[0201] (Purification Process) The solution containing the obtained peptide-containing polyvinyl acetal resin (A) was diluted 100-fold with DMF and washed by dropping it onto a column packed with ion exchange resin (manufactured by Organo) at a rate of 0.3 mL / min. The washed solution was vacuum-dried at 60°C for 3 hours to obtain a dry product. The obtained dry product was dissolved in butanol (alcohol solvent), and 5 parts by weight of acetic acid (pH adjuster) was added to 100 parts by weight of butanol to obtain a coating solution containing peptide-containing polyvinyl acetal resin (A) and butanol. The content of peptide-containing polyvinyl acetal resin (A) in the coating solution was 0.1% by weight.

[0202] (Preparation of cell culture substrate) 30 μL of the obtained coating solution was coated onto each well of a 12-well plate by cast coating, and then vacuum-dried at 60°C for 3 hours to remove the alcohol solvent. In this way, a cell culture substrate (cell culture container) was obtained in which a cell scaffold material (resin film), which is a dried layer of the coating solution, was placed on the bottom surface of each well of the 12-well plate.

[0203] (Example 2) Except that the amount of the first peptide used was changed to 6 parts by weight and the amount of the second peptide used was changed to 6 parts by weight in the preparation step, a cell scaffold material and a cell culture substrate were obtained in the same manner as in Example 1.

[0204] (Example 3) Except that the amount of the first peptide used in the preparation step was changed to 11.3 parts by weight and the amount of the second peptide used to 0.8 parts by weight, a cell scaffold material and a cell culture substrate were obtained in the same manner as in Example 1.

[0205] (Example 4) Except that the amount of the first peptide used was changed to 10.5 parts by weight and the amount of the second peptide used to 1.5 parts by weight in the preparation step, a cell scaffold material and a cell culture substrate were obtained in the same manner as in Example 1.

[0206] (Example 5) Except that the amount of the first peptide used was changed to 7.5 parts by weight and the amount of the second peptide used to 4.5 parts by weight in the preparation step, a cell scaffold material and a cell culture substrate were obtained in the same manner as in Example 1.

[0207] (Example 6) Except that the amount of the first peptide used was changed to 4.5 parts by weight and the amount of the second peptide used to 7.5 parts by weight in the preparation step, a cell scaffold material and a cell culture substrate were obtained in the same manner as in Example 1.

[0208] (Example 7) Except that the amount of the first peptide used was changed to 1.5 parts by weight and the amount of the second peptide used to 10.5 parts by weight in the preparation step, a cell scaffold material and a cell culture substrate were obtained in the same manner as in Example 1.

[0209] (Example 8) Except that the amount of the first peptide used was changed to 0.8 parts by weight and the amount of the second peptide used to 11.3 parts by weight in the preparation step, a cell scaffold material and a cell culture substrate were obtained in the same manner as in Example 1.

[0210] (Example 9) A cell scaffold material and a cell culture substrate were obtained in the same manner as in Example 1, except that a peptide having the amino acid sequence Arg-Gly-Asp-Ser (4 amino acid residues, RGDS) was used as the first peptide.

[0211] (Example 10) A cell scaffold material and a cell culture substrate were obtained in the same manner as in Example 1, except that a peptide having the amino acid sequence Gly-Arg-Gly-Asp-Ser (5 amino acid residues, GRGDS) was used as the first peptide.

[0212] (Example 11) A cell scaffold material and a cell culture substrate were obtained in the same manner as in Example 1, except that a peptide having the amino acid sequence Leu-Asp-Val (3 amino acid residues, LDV) was used as the second peptide.

[0213] (Example 12) A cell scaffold material and a cell culture substrate were obtained in the same manner as in Example 1, except that the amount of the first peptide used was changed to 15.0 parts by weight and the amount of the second peptide used was changed to 15.0 parts by weight in the preparation step.

[0214] (Example 13) A cell scaffold material and a cell culture substrate were obtained in the same manner as in Example 1, except that the amount of the first peptide used in the preparation step was changed to 25.5 parts by weight and the amount of the second peptide used was changed to 25.5 parts by weight.

[0215] (Example 14) A cell scaffold material and a cell culture substrate were obtained in the same manner as in Example 1, except that the amount of the first peptide used in the preparation step was changed to 30.8 parts by weight and the amount of the second peptide used was changed to 30.8 parts by weight.

[0216] (Example 15) A cell scaffold material and a cell culture substrate were obtained in the same manner as in Example 1, except that the amount of the first peptide used was changed to 0.1 parts by weight and the amount of the second peptide used was changed to 0.1 parts by weight in the preparation step.

[0217] (Example 16) An acrylic monomer solution was obtained by dissolving 75 parts by weight of N-isopropylacrylamide and 25 parts by weight of butyl methacrylate in 300 parts by weight of tetrahydrofuran. Two parts by weight of Irgacure 184 (manufactured by BASF) were dissolved in the obtained acrylic monomer solution and coated onto a PET film. The coated object was exposed to light at a wavelength of 365 nm with an integrated light intensity of 2000 mJ / cm using an I-Graphics UV conveyor device "ECS301G1" at 25°C. ２ An acrylic resin solution was obtained by irradiation. The obtained acrylic resin solution was vacuum-dried at 80°C for 3 hours to obtain acrylic resin. Cell scaffold material and cell culture substrate were obtained in the same manner as in Example 1, except that the obtained acrylic resin was used instead of polyvinyl acetal resin.

[0218] (Example 17) An acrylic resin was obtained in the same manner as in Example 16, except that 90 parts by weight of methoxyethyl acrylate and 10 parts by weight of butyl methacrylate were used instead of 75 parts by weight of N-isopropylacrylamide and 25 parts by weight of butyl methacrylate. Furthermore, a cell scaffold material and a cell culture substrate were obtained in the same manner as in Example 16, except that this acrylic resin was used.

[0219] (Example 18) An acrylic resin was obtained in the same manner as in Example 16, except that 75 parts by weight of methoxyethyl acrylate and 25 parts by weight of butyl methacrylate were used instead of 75 parts by weight of N-isopropyl acrylamide and 25 parts by weight of butyl methacrylate. Furthermore, a cell scaffold material and a cell culture substrate were obtained in the same manner as in Example 16, except that this acrylic resin was used.

[0220] (Example 19) An acrylic resin was obtained in the same manner as in Example 16, except that 2 parts by weight of butyl methacrylate and 98 parts by weight of ethyl acrylate were used instead of 75 parts by weight of N-isopropylacrylamide and 25 parts by weight of butyl methacrylate. Furthermore, a cell scaffold material and a cell culture substrate were obtained in the same manner as in Example 16, except that this acrylic resin was used.

[0221] (Example 20) (Preparation of coating solution X) A polyvinyl acetal resin X (polyvinyl acetal resin bonded with acrylic acid and butyl acrylate) prepared in Example 1 was prepared. DMF was prepared as the first solvent. A cyclic peptide having the amino acid sequence Arg-Gly-Asp-Phe-Lys (5 amino acid residues, Arg and Lys bond to form a cyclic skeleton, Phe is the D-isomer, c-RGDfK) was prepared as the first peptide. 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride was prepared as the condensing agent. 50 parts by weight of polyvinyl acetal resin X and 3 parts by weight of the first peptide were mixed with 1000 parts by weight of the first solvent to prepare the first solution. A second solution was prepared by mixing 10 parts by weight of the condensing agent with 10 parts by weight of the first solvent. The first solution and the second solution were mixed to prepare a solution containing polyvinyl acetal resin X, the first peptide, and a condensing agent.

[0222] The obtained solution was reacted at 40°C for 2 hours to dehydrate and condense the carboxyl groups in the structural units derived from acrylic acid of polyvinyl acetal resin X with the amino groups of Lys in the first peptide, thereby obtaining a solution containing peptide-containing polyvinyl acetal resin (1).

[0223] The solution containing the obtained peptide-containing polyvinyl acetal resin (1) was diluted 100-fold with DMF and washed by dropping it dropwise into a column packed with ion exchange resin (manufactured by Organo) at a rate of 0.3 mL / min. The washed solution was vacuum-dried at 60°C for 3 hours to obtain a dry product. The obtained dry product was dissolved in butanol (alcohol solvent), and 5 parts by weight of acetic acid (pH adjuster) was added to 100 parts by weight of butanol to obtain a coating solution X containing peptide-containing polyvinyl acetal resin (1) and butanol. The content of peptide-containing polyvinyl acetal resin (1) in the coating solution X was 0.1% by weight.

[0224] (Preparation of coating solution Y) A polyvinyl acetal resin X (polyvinyl acetal resin bonded with acrylic acid and butyl acrylate) prepared in Example 1 was prepared. DMF was prepared as the second solvent. A peptide having the amino acid sequence Glu-Ile-Leu-Asp-Val (5 amino acid residues, EILDV) was prepared as the second peptide. 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride was prepared as the condensing agent. 50 parts by weight of polyvinyl acetal resin X and 3 parts by weight of the second peptide were mixed with 1000 parts by weight of the second solvent to prepare the first solution. Also, 10 parts by weight of the condensing agent were mixed with 1000 parts by weight of the second solvent to prepare the second solution. The first solution and the second solution were mixed to prepare a solution containing polyvinyl acetal resin X, the second peptide and the condensing agent.

[0225] The obtained solution was reacted at 40°C for 2 hours to dehydrate and condense the carboxyl groups in the structural units derived from acrylic acid of polyvinyl acetal resin X with the amino groups of Glu in the second peptide, thereby obtaining a solution containing peptide-containing polyvinyl acetal resin (2).

[0226] The solution containing the obtained peptide-containing polyvinyl acetal resin (2) was diluted 100-fold with DMF and washed by dropping it dropwise into a column packed with ion exchange resin (manufactured by Organo) at a rate of 0.3 mL / min. The washed solution was vacuum-dried at 60°C for 3 hours to obtain a dry product. The obtained dry product was dissolved in butanol (alcohol solvent), and 5 parts by weight of acetic acid (pH adjuster) was added to 100 parts by weight of butanol to obtain a coating solution Y containing peptide-containing polyvinyl acetal resin (2) and butanol. The content of peptide-containing polyvinyl acetal resin (2) in the coating solution Y was 0.1% by weight.

[0227] (Preparation of cell scaffold material and cell culture substrate) Coating solution X was mixed with coating solution Y, 50 parts by weight to obtain coating solution Z. Except for using coating solution Z, a cell scaffold material and a cell culture substrate were obtained in the same manner as in Example 1.

[0228] (Example 21) A cell scaffold material and a cell culture substrate were obtained in the same manner as in Example 20, except that the amount of the first peptide used in the preparation step of coating solution X was changed to 11.3 parts by weight, and the amount of the second peptide used in the preparation step of coating solution Y was changed to 0.8 parts by weight.

[0229] (Example 22) A cell scaffold material and a cell culture substrate were obtained in the same manner as in Example 20, except that the amount of the first peptide used in the preparation step of coating solution X was changed to 0.8 parts by weight, and the amount of the second peptide used in the preparation step of coating solution Y was changed to 11.3 parts by weight.

[0230] (Comparative Example 1) A cell scaffold material and a cell culture substrate were obtained in the same manner as in Example 1, except that the first peptide was not used.

[0231] (Comparative Example 2) A cell scaffold material and a cell culture substrate were obtained in the same manner as in Example 1, except that the second peptide was not used.

[0232] (Comparative Example 3) Except for changing the amount of n-butyraldehyde (n-BA) added in the second step of preparing the polyvinyl acetal resin X from 148 g to 89 g, a cell scaffold material and a cell culture substrate were obtained in the same manner as in Example 1.

[0233] (Comparative Example 4) A polyethyl acrylate resin was obtained by mixing 100 parts by weight of ethyl acrylate, 75 parts by weight of ethyl acetate, and 0.5 parts by weight of azobisisobutyronitrile, and polymerization was carried out at 65°C for 8 hours under a nitrogen atmosphere. A cell scaffold material and a cell culture substrate were obtained in the same manner as in Example 1, except that the obtained polyethyl acrylate resin was used instead of the polyvinyl acetal resin.

[0234] (Comparative Example 5) A cell scaffold material and a cell culture substrate were obtained in the same manner as in Comparative Example 4, except that 100 parts by weight of butyl methacrylate were used instead of 100 parts by weight of ethyl acrylate.

[0235] (Evaluation) (1) (Surface Free Energy) The surface free energy of the obtained cell scaffold material (resin film) was measured using a contact angle meter (Kyowa Interface Chemical Co., Ltd., DMo-701). The contact angle of pure water was obtained by dropping 1 μL of pure water onto the resin film and taking a picture of the droplet after 10 seconds. The contact angle of diiodomethane was obtained by dropping 1 μL of diiodomethane onto the resin film and taking a picture of the droplet after 10 seconds. From the obtained contact angles, the dispersion component γ of the surface free energy was calculated using the Kaelble-Uy theoretical formula. ｄ and dipole component γ ｐ The result was calculated.

[0236] (2) Preparation of medium A for maintaining memory T cells and T cell proliferation performance: IL-2 (Myltenyi Biotec) was added to TexMACS medium (Myltenyi Biotec) to a concentration of 10 ng / mL. Then, ImmunoCult Human CD3 / CD28 / CD2 T cell activator (Stemcell Technologies) was added to a concentration of 25 μL / mL to obtain medium A.

[0237] Preparation of medium B: Medium B was prepared by adding IL-2 to TexMACS medium to a concentration of 10 ng / mL.

[0238] Preparation of RetroNectin-coated plate (control 1): RetroNectin (Takara Bio Inc.) was diluted with PBS to a concentration of 25 μg / mL, and 1 mL / well was added to a 12-well TissueCulture plate. The plate was left to stand overnight at 4°C or at room temperature for 2 hours. Before use, the solution in the wells was removed, and the wells were washed by adding and removing 1 mL / well of PBS. The wells were washed twice.

[0239] Preparation of an uncoated plate (control 2): ​​The 12-well TissueCulture Plate itself was used as the uncoated plate.

[0240] T cell culture: The frozen tubes containing T cells (LONZA) were warmed and thawed in a 37°C constant temperature water bath. A 15 mL tube containing 10 mL of TexMACS warmed to 37°C was prepared. The thawed T cells were added to this 15 mL tube. Then, centrifugation was performed at 300 × g for 10 minutes, the supernatant was removed, and the T cell pellet was suspended in 2 mL of medium A. After counting the number of cells with TC20 (BioRAD), 1 × 10¹⁶ cells were added to medium A. ６ A cell suspension was prepared at a concentration of cells / mL.

[0241] Prepared cell culture substrates (cell culture vessels equipped with cell scaffold material containing peptide-containing resin, 12-well plates), a RetroNectin-coated plate (control 1), and an uncoated plate (control 2). One mL of the cell suspension was seeded into each well of these plates. Afterward, the plates were shaken to homogenize the cells, and then CO2 was added. ２ Incubator at 37°C and 5% CO2 ２ Culturing was started under the following conditions. Two days after sowing, 1 mL / well of medium B was added. Five days after sowing, 2 mL / well of medium B was added.

[0242] Seven days after seeding, the cells were pipetted out of the wells, detached, suspended, and collected in tubes. 400 μL of medium B was added to the collected wells and collected, then added to the tubes. Next, 0.5 mL / well of TexMACS was added to the empty wells. The tubes containing the collected cells were centrifuged at 300 × g for 10 minutes, the supernatant was removed, and the cells were suspended in 2 mL of medium B. After counting the number of cells in the suspension, 2 × 10⁶ cells were added to medium B. ５ Prepared at cells / mL. After removing the culture medium from the original wells, 1 mL of the cell suspension recovered from the corresponding well was immediately added to each well. After shaking the plate to homogenize the cells, CO2 was added. ２ Incubator at 37°C and 5% CO2 ２ The culture was restarted under these conditions.

[0243] Two days after sowing, 1 mL / well of medium B was added. Furthermore, 2 mL / well of medium B was added on the fifth day after sowing.

[0244] Seven days after seeding (14th day of culture), cells were pipetted from the wells to detach and suspend, then collected in tubes. 400 μL of medium B was added to the collected wells and collected, then added to the tubes. The tubes containing the collected cells were centrifuged at 300 × g for 10 minutes, the supernatant was removed, and the cells were suspended in 2 mL of medium A. After counting the number of cells in the suspension, 2 × 10⁶ cells were added to medium A. ５ Prepared at a concentration of 1 / mL.

[0245] New cell culture substrates were prepared (cell culture vessels equipped with cell scaffold material containing peptide-containing resin, 12-well plates), a RetroNectin-coated plate (control 1), and an uncoated plate (control 2). 1 mL of cell suspension recovered from the corresponding well was added to each well of these plates. After shaking the plates to homogenize the cells, CO2 was added. ２ Incubator at 37°C and 5% CO2 ２ The culture was restarted under the following conditions. Two days after sowing, 1 mL / well of medium B was added. Five days after sowing, 2 mL / well of medium B was added.

[0246] Seven days after seeding (21 days of culture), cells were pipetted from the wells to detach and suspend, then collected in tubes. 400 μL of medium B was added to the collected wells and collected, then added to the tubes. After collecting the cells from the wells, 0.5 mL / well of TexMACS was added to an empty well. The tubes containing the collected cells were centrifuged at 300 × g for 10 minutes, the supernatant was removed, and the cells were suspended in 2 mL of medium B. After counting the number of cells in the suspension, 2 × 10⁶ cells were added in medium B. ５ Prepared at cells / mL. After removing the culture medium from the original wells, 1 mL of the cell suspension recovered from the corresponding well was immediately added to each well. After shaking the plate to homogenize the cells, CO2 was added. ２ Incubator at 37°C and 5% CO2 ２ The culture was restarted under the following conditions. Two days after sowing, medium B was added at a rate of 1 mL / well. On the fifth day after sowing, medium B was added at a rate of 2 mL / well.

[0247] Seven days after seeding (28 days into culture), cells were detached from the wells by pipetting, suspended, and collected in tubes. 400 μL of medium B was added to the collected wells and collected, then added to the tubes. The tubes containing the collected cells were centrifuged at 300 × g for 10 minutes, the supernatant was removed, and the cells were suspended in 2 mL of TexMACS. The number of cells in the suspension was counted.

[0248] However, the evaluation of memory T cell maintenance was performed on day 21 of the culture process described above. At that time, on Day 21, cells were detached from the wells by pipetting, suspended, and collected in a tube. 400 μL of medium B was added to the wells after collection and collected, and added to the tube described above. The tube containing the collected cells was centrifuged at 300 × g for 10 minutes, the supernatant was removed, and the cells were suspended in 2 mL of medium B. After counting the number of cells in the suspension, 5 × 10⁴ cells were counted. ５ ~1 x 10 ６ Individual cells were collected in separate tubes and used as samples for flow cytometry.

[0249] Evaluation of memory T cell maintenance (memory T cell markers): 1 × 10⁶ cells obtained from a cell suspension cultured for 21 days. ５Cells were collected in 1.5 mL tubes. These tubes were centrifuged at 300 × g for 5 minutes, the supernatant was removed, and 300 μL of Fix Buffer I (BD Japan), warmed at 37°C, was added. After incubation at 37°C for 10 minutes, the cells were centrifuged at 300 × g for 5 minutes, and the supernatant was removed. 1 mL of Stain Buffer was added, and the cells were centrifuged at 300 × g for 5 minutes, the supernatant was removed, and the cells were suspended in 50 μL of Stain Buffer (BD Japan). After suspension, 5 μL of HumanBD Fc Block (BD Japan) was added and mixed, and the cells were allowed to stand at room temperature for 10 minutes. Subsequently, antibodies (fluorescent dye-conjugated) specific to the human memory marker CD45RA and the fatigue marker PD-1 were added, and the cells were stained in a refrigerator under light protection for 30 minutes. No antibody was added to the negative control. After staining for 30 minutes, 1 mL of Stain Buffer was added and mixed. Then, centrifugation was performed at 300 × g for 5 minutes. After centrifugation, the supernatant was removed without disturbing the cell pellet, and 300 μL of Stain Buffer was added again to resuspend the cells. The cell suspension was measured using a flow cytometry system (BD Japan) to determine the percentage of CD45RA-positive and PD-1-negative cells.

[0250] The percentage of CD45RA-positive and PD-1-negative cells in cell samples cultured on a RetroNectin-coated plate (control 1) was defined as "Percentage A," and the percentage of CD45RA-positive and PD-1-negative cells in cell samples cultured on a cell culture substrate (a cell culture vessel equipped with a cell scaffold material containing peptide-containing resin) was defined as "Percentage B." Percentage X (%) was then calculated by "Percentage B ÷ Percentage A × 100."

[0251] [Criteria for determining the maintenance of memory T cells] ○○○: Percentage X is 150% or more ○○: Percentage X is 125% or more but less than 150% ○: Percentage X is 100% or more but less than 125% ×: Percentage X is less than 100%

[0252] Evaluation of T cell proliferation: T cell proliferation was compared by calculating the proliferation rate R(0-28) from the start of culture to day 28 of culture. When the number of cells seeded on the day the cell stock was thawed and culture started is defined as Day 0, and the number of cells collected on day 7 (Day 7) is defined as N-IN(0), the proliferation rate R(0-7) at Day 7 is expressed as follows. Note that these cell counts refer to living cells.

[0253] R(0-7)=N-OUT(7) / N-IN(0)

[0254] When the number of cells seeded on Day 7 is N-IN(7) and the number of cells collected on Day 14 is N-OUT(14), the proliferation rate R(7-14) from Day 7 to Day 14 is expressed as follows.

[0255] R(7-14)=N-OUT(14) / N-IN(7)

[0256] The growth rate R(0-14) from the start of culture (Day 0) to Day 14 is expressed as follows:

[0257] R(0-14)=R(0-7)×R(7-14)

[0258] As described above, the proliferation rate during the culture period was calculated and the proliferative capacity of the cells was compared. The viability was obtained using an automated cell counting device TC20 (BioRad) with the cell suspension obtained at the end of the culture period.

[0259] The growth rate during the culture period, i.e., the growth rate R(0-28) from the start of culture to Day 28, was determined. The growth rate R(0-28) for cell samples cultured on a RetroNectin coated plate (control 1) was defined as "growth rate RA," and the growth rate R(0-28) for cell samples cultured on a cell culture substrate (a cell culture vessel equipped with a cell scaffold material containing peptide-containing resin) was defined as "growth rate RB." The growth rate X (%) was then calculated using the formula "growth rate RB ÷ growth rate RA × 100."

[0260] [Criteria for determining T cell proliferation] ○○○: Proliferation rate X is 200% or more ○○: Proliferation rate X is 100% or more but less than 200% ○: Proliferation rate X is 50% or more but less than 100% ×: Proliferation rate X is less than 50%

[0261] (3) The solubility of peptide-containing resin in the coating solution was quantified using a dynamic light scattering (DLS) measuring device ("NANOTRAC WAVE I" manufactured by Microtrac-Bell). First, SetZ (SZ) was performed with a solution of ethanol (95 wt%) / acetic acid (5 wt%). The coating solution was filled into the measuring device and processed in the order of SL (Sample Load) and Run. The average value of the results of a total of three measurements was taken as the Loading Index. A smaller Loading Index indicates higher transparency of the coating solution and better solubility of the peptide-containing resin in the coating solution.

[0262] [Criteria for determining solubility] A: Loading Index is 0.1 or less B: Loading Index is greater than 0.1 and 0.5 or less C: Loading Index is greater than 0.5

[0263] The configuration and results are shown in Tables 1 to 17 below.

[0264]

[0265]

[0266]

[0267]

[0268]

[0269]

[0270]

[0271]

[0272]

[0273]

[0274]

[0275]

[0276]

[0277]

[0278]

[0279]

[0280]

[0281] 1...Cell culture substrate 2...Substrate body 2a...First surface 3...Cell scaffold material 10...Peptide-containing resin (A) 10a...Peptide-containing resin (1) 10b...Peptide-containing resin (2) 11, 11a, 11b...Synthetic resin part 12, 12a, 12b...Linker part 13, 13a...First peptide part 14, 14b...Second peptide part 15...Peptide part

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