Compositions and methods useful for culturing differentiable cells
First Claim
1. A method of expanding pluripotent stem cells, said method comprisinga) culturing the pluripotent stem cells on a solid surface in an adherent culture in a cell medium, the cell medium comprising a basal salt nutrient solution and an ErbB3 ligand, where the cell medium is essentially free of serum, wherein the pluripotent stem cells can give rise to all three germ layers and wherein the ErbB3 ligand binds to an ErbB3 receptor, which in turn dimerizes to an ErbB2 receptor to form an ErbB2/ErbB3 heterodimer, thereby activating the tyrosine kinase activity of the ErbB2 receptor in the ErbB2/ErbB3 heterodimer;
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Abstract
The present invention relates to cell culture methods and compositions that are essentially serum-free and comprise a basal salt nutrient solution and an ErbB3 ligand.
40 Citations
33 Claims
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1. A method of expanding pluripotent stem cells, said method comprising
a) culturing the pluripotent stem cells on a solid surface in an adherent culture in a cell medium, the cell medium comprising a basal salt nutrient solution and an ErbB3 ligand, where the cell medium is essentially free of serum, wherein the pluripotent stem cells can give rise to all three germ layers and wherein the ErbB3 ligand binds to an ErbB3 receptor, which in turn dimerizes to an ErbB2 receptor to form an ErbB2/ErbB3 heterodimer, thereby activating the tyrosine kinase activity of the ErbB2 receptor in the ErbB2/ErbB3 heterodimer; - and
b) maintaining the pluripotent stem cells in the adherent culture in the cell medium and in appropriate culture conditions for more than one day, wherein the cell medium is changed periodically, wherein the pluripotent stem cells proliferate and do not differentiate. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18)
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19. A method of expanding primate embryonic stem cells, the method comprising:
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a) culturing the primate embryonic stem cells on a solid surface in an adherent culture in a cell medium, the cell medium comprising a basal salt nutrient solution and an ErbB3 ligand, where the cell medium is essentially free of serum, wherein the primate embryonic stem cells can give rise to all three germ layers and wherein the ErbB3 ligand binds to an ErbB3 receptor, which in turn dimerizes to an ErbB2 receptor to form an ErbB2/ErbB3 heterodimer, thereby activating the tyrosine kinase activity of the ErbB2 receptor in the ErbB2/ErbB3 heterodimer; and b) maintaining the primate embryonic stem cells in the adherent culture in the cell medium and in appropriate culture conditions for more than one day, wherein the cell medium is changed periodically, wherein the primate embryonic stem cells proliferate and do not differentiate. - View Dependent Claims (20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33)
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Specification