Cell culture member and method for modifying surface thereof

Patent metadata

Field	Value
Publication number	US-2024052299-A1
Application number	US-202118259237-A
Country	US
Kind code	A1
Filing date	Dec 23, 2021
Priority date	Dec 25, 2020
Publication date	Feb 15, 2024
Grant date	—

Abstract

Provided are a cell culture member having excellent cell culture performance and its long-term stability, and a method for modifying the surface thereof. The cell culture member according to the present invention is a cell culture member having at least a holding region that holds an adherent cell and contains a polymer compound, wherein at least a part of the holding region is a surface-modified region in which a fluorine atom is directly chemically bonded to a part of carbon atoms and/or silicon atoms constituting the polymer compound, and the present invention can provide a cell culture member that improves adhesiveness of the adherent cell to the surface-modified region, suppresses deterioration over time of adhesiveness, and is excellent in cell culture performance and its long-term stability.

First claim

1 . A cell culture member having at least a holding region that holds an adherent cell and contains a polymer compound, wherein at least a part of the holding region is a surface-modified region in which a fluorine atom is directly chemically bonded to a part of carbon atoms and/or silicon atoms constituting the polymer compound.

2 . The cell culture member according to claim 1 , wherein a peak value of binding energy of the fluorine atom, as measured by X-ray photoelectron spectroscopy, is in a range of 680 eV to 690 eV.

3 . The cell culture member according to claim 1 , wherein
a surface-modifying group is directly chemically bonded to a part of other carbon atoms and/or other silicon atoms constituting the polymer compound in the surface-modified region, the surface-modifying group is at least one selected from a group consisting of an —OR 1 group; a —COOR 2 group; a —COR 3 group; a hydrocarbon group; a silyl group; a hydrocarbon group having at least one of a hetero atom, a halogen atom, or an unsaturated bond; a silyl group having at least one of a hetero atom, a halogen atom, or an unsaturated bond; a cyano group; a nitro group; a nitroso group; a phosphate group; a sulfonyl group; a thiol group; a thionyl group; and a halogen atom excluding a fluorine atom, R 1 and R 2 are each independently a hydrogen atom; a metal atom; a hydrocarbon group; a silyl group; a hydrocarbon group having at least one of a hetero atom, a halogen atom, or an unsaturated bond; or a silyl group having at least one of a hetero atom, a halogen atom, or an unsaturated bond, and R 3 is a hydrocarbon group; a hydrocarbon group having at least one of a hetero atom or an unsaturated bond; or a silyl group having at least one of a hetero atom or an unsaturated bond.

4 . The cell culture member according to wherein the polymer compound is at least one polymer selected from a group consisting of polyvinyl chloride, polystyrene, polyethylene, polypropylene, polyvinyl acetate, polyurethane, cyclic polyolefin, polyether ether ketone, polyimide, polyamide imide, polycarbonate, polymethyl methacrylate, polyethylene terephthalate, acrylonitrile-butadiene-styrene, polyacrylonitrile, polyamide, polyvinyl alcohol, polyolefin, and a silicon-containing polymer compound.

5 . A method for modifying a surface of a cell culture member having a holding region that holds an adherent cell and contains a polymer compound,
the method comprising a step of bringing a first treatment gas containing: a gas containing a fluorine atom; and an inert gas as an optional component into contact with at least a part of the holding region, and forming a surface-modified region in which the fluorine atom is directly chemically bonded to a part of carbon atoms and/or silicon atoms constituting the polymer compound.

6 . The method for modifying a surface of a cell culture member according to claim 5 , wherein a peak value of binding energy of the fluorine atom in the surface-modified region, as measured by X-ray photoelectron spectroscopy, is in a range of 680 eV to 690 eV.

7 . The method for modifying a surface of a cell culture member according to claim 5 , wherein
in the step, a surface-modifying group is directly chemically bonded to a part of other carbon atoms and/or other silicon atoms constituting the polymer compound by using a gas further containing a gas containing an oxygen atom as the first treatment gas, the surface-modifying group is at least one selected from a group consisting of an —OR 1 group; a —COOR 2 group; a —COR 3 group; a hydrocarbon group; a silyl group; a hydrocarbon group having at least one of a hetero atom, a halogen atom, or an unsaturated bond; a silyl group having at least one of a hetero atom, a halogen atom, or an unsaturated bond; a nitro group; a nitroso group; a phosphate group; a sulfonyl group; a thionyl group; and a halogen atom excluding a fluorine atom, R 1 and R 2 are each independently a hydrogen atom; a metal atom; a hydrocarbon group; a silyl group; a hydrocarbon group having at least one of a hetero atom, a halogen atom, or an unsaturated bond; or a silyl group having at least one of a hetero atom, a halogen atom, or an unsaturated bond, and R 3 is a hydrocarbon group; a hydrocarbon group having at least one of a hetero atom or an unsaturated bond; or a silyl group having at least one of a hetero atom or an unsaturated bond.

8 . The method for modifying a surface of a cell culture member according to claim 5 , further comprising a step of bringing a second treatment gas containing another gas containing an oxygen atom into contact with at least a part of the holding region which has been brought into contact with the first treatment gas, and causing a surface-modifying group to be directly chemically bonded to a part of other carbon atoms and/or other silicon atoms constituting the polymer compound,
wherein the surface-modifying group is at least one selected from a group consisting of an —OR 1 group; a —COOR 2 group; a —COR 3 group; a hydrocarbon group; a silyl group; a hydrocarbon group having at least one of a hetero atom, a halogen atom, or an unsaturated bond; a silyl group having at least one of a hetero atom, a halogen atom, or an unsaturated bond; a nitro group; a nitroso group; a phosphate group; a sulfonyl group; a thionyl group; and a halogen atom excluding a fluorine atom, R 1 and R 2 are each independently a hydrogen atom; a metal atom; a hydrocarbon group; a silyl group; a hydrocarbon group having at least one of a hetero atom, a halogen atom, or an unsaturated bond; or a silyl group having at least one of a hetero atom, a halogen atom, or an unsaturated bond, and R 3 is a hydrocarbon group; a hydrocarbon group having at least one of a hetero atom or an unsaturated bond; or a silyl group having at least one of a hetero atom or an unsaturated bond.

9 . The method for modifying a surface of a cell culture member according to claim 7 , further comprising a step of bringing the fluorine atom directly chemically bonded to the part of the carbon atoms and/or the silicon atoms and/or the surface-modifying group directly chemically bonded to the part of the other carbon atoms and/or the other silicon atoms into contact with a treatment compound that reacts with the fluorine atom and/or the surface-modifying group, and substituting the fluorine atom and/or the surface-modifying group with a surface-modifying group derived from the treatment compound.

10 . The method for modifying a surface of a cell culture member according to claim 5 , wherein the polymer compound is at least one polymer selected from a group consisting of polyvinyl chloride, polystyrene, polyethylene, polypropylene, polyvinyl acetate, polyurethane, cyclic polyolefin, polyether ether ketone, polyimide, polyamide imide, polycarbonate, polymethyl methacrylate, polyethylene terephthalate, acrylonitrile-butadiene-styrene, polyacrylonitrile, polyamide, polyvinyl alcohol, polyolefin, and a silicon-containing polymer compound.

11 . The method for modifying a surface of a cell culture member according to claim 7 , further comprising a step of bringing the fluorine atom directly chemically bonded to the part of the carbon atoms and/or the silicon atoms and/or the surface-modifying group directly chemically bonded to the part of the other carbon atoms and/or the other silicon atoms into contact with a treatment compound that reacts with the fluorine atom and/or the surface-modifying group, and substituting the fluorine atom and/or the surface-modifying group with a surface-modifying group derived from the treatment compound.

Assignees

• Univ Shinshu
• Stella Chemifa Corp

Inventors

• Saito Naoto
• Uemura Takeshi
• HISANO Kazutoshi
• Sato Yoshinori
• Yamamoto Masashi
• Nii Keiichi
• Nishida Tetsuo

Classifications

• C12N5/0068 Primary

  General culture methods using substrates (for specific animal cell type C12N5/06) · CPC title

• C12M25/04 Primary

  in combination with well or multiwell plates, i.e. culture inserts · CPC title

• C08J7/12

  Chemical modification · CPC title

• C12N2509/00

  Methods for the dissociation of cells, e.g. specific use of enzymes · CPC title

• C12N2533/30

  Synthetic polymers · CPC title