Cell culture substrate, and cell culturing method using the substrate and method for inducing differentiation of pluripotent stem cells using the substrate
First Claim
1. A cell culture substrate, which is used for culturing for induction into neural progenitor cells or neural cells, characterized in that, onto the surface, at least two proteins are immobilized or coated, wherein said at least two proteins comprise:
- a first protein which is selected from the group consisting of a fusion protein comprising an entire or partial region of N-cadherin, and a fusion protein comprising an entire or partial region of a protein homologous to N-cadherin; and
a second protein which is selected from the group consisting of a protein belonging to cadherin family other than N-cadherin, a fusion protein comprising at least one extracellular (EC) domain selected from EC1 domain, EC2 domain, EC3 domain, EC4 domain and EC5 domain of a protein belonging to cadherin family other than N-cadherin, and a fusion protein comprising at least one extracellular (EC) domain selected from EC1 domain, EC2 domain, EC3 domain, EC4 domain and EC5 domain of a protein homologous to a protein belonging to cadherin family other than N-cadherin, wherein said protein belonging to cadherin family other than N-Cadherin is E-cadherin wherein the concentration of the first protein is 3-1000 μ
g/mL and the concentration of the second protein is 0.01-1000 μ
g/mL;
ac4 wherein the substrate comprises polystyrene, polyethylene or polypropylene; and
wherein said protein homologous to N-cadherin is a protein which comprises one or more of the EC1 domain, EC2 domain, EC3 domain, EC4 domain and EC5 domain, and which has hemophilic binding ability with N-cadherin.
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Abstract
[Problem] To provide a cell culture substrate, and a cell culturing method using the substrate and a method for inducing differentiation of pluripotent stem cells using the substrate, which allow culturing of pluripotent stem cells and allow differentiation of pluripotent stem cells into a specified cell species, particularly neural and neural progenitor cells, at a high purity.
[Means for Solution] A cell culture substrate, characterized in that, onto the surface, one or more selected from the group consisting of N-cadherin, a fusion protein comprising an entire or partial region of N-cadherin, and a fusion protein comprising an entire or partial region of a protein homologous to N-cadherin are immobilized or coated.
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Citations
15 Claims
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1. A cell culture substrate, which is used for culturing for induction into neural progenitor cells or neural cells, characterized in that, onto the surface, at least two proteins are immobilized or coated, wherein said at least two proteins comprise:
- a first protein which is selected from the group consisting of a fusion protein comprising an entire or partial region of N-cadherin, and a fusion protein comprising an entire or partial region of a protein homologous to N-cadherin; and
a second protein which is selected from the group consisting of a protein belonging to cadherin family other than N-cadherin, a fusion protein comprising at least one extracellular (EC) domain selected from EC1 domain, EC2 domain, EC3 domain, EC4 domain and EC5 domain of a protein belonging to cadherin family other than N-cadherin, and a fusion protein comprising at least one extracellular (EC) domain selected from EC1 domain, EC2 domain, EC3 domain, EC4 domain and EC5 domain of a protein homologous to a protein belonging to cadherin family other than N-cadherin, wherein said protein belonging to cadherin family other than N-Cadherin is E-cadherin wherein the concentration of the first protein is 3-1000 μ
g/mL and the concentration of the second protein is 0.01-1000 μ
g/mL;
ac4 wherein the substrate comprises polystyrene, polyethylene or polypropylene; and
wherein said protein homologous to N-cadherin is a protein which comprises one or more of the EC1 domain, EC2 domain, EC3 domain, EC4 domain and EC5 domain, and which has hemophilic binding ability with N-cadherin. - View Dependent Claims (2, 3, 4, 5, 7, 8, 9, 10, 11, 12, 13, 14, 15)
- a first protein which is selected from the group consisting of a fusion protein comprising an entire or partial region of N-cadherin, and a fusion protein comprising an entire or partial region of a protein homologous to N-cadherin; and
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6. A cell culture substrate, characterized in that, onto the surface, two or more selected from the group consisting of a protein belonging to cadherin family and a fusion protein comprising an entire or partial region of a protein belonging to cadherin family are immobilized or coated.
Specification