Methods for inducing differentiation of pluripotent cells
First Claim
1. A method for inducing differentiation of pluripotent cells comprising the following steps (a) and (b):
- (a) culturing the pluripotent cells in a medium comprising any one of the following growth factors (i) to (iii);
(i) acidic fibroblast growth factor, fibroblast growth factor 4, and hepatocyte growth factor;
(ii) acidic fibroblast growth factor, and growth factor(s) selected from activin A, epidermal growth factor, and β
-nerve growth factor; and
(iii) fibroblast growth factor 4, and growth factor(s) selected from activin A and hepatocyte growth factor; and
, (b) culturing the cell cultured in step (a) in a medium comprising oncostatin M.
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Abstract
The object of the present invention is to provide methods for identifying factors involved in inducing the differentiation of pluripotent cells, and methods for using those factors.
Using cDNA microarray technology, variations in gene expression between placebo-treated and CCl4-treated mice livers were analyzed 24 hours after treatment, and several growth factors were selected. The effects of matrix and growth factors on hepatocyte differentiation rates of ES cells were investigated. As a result, the present inventors succeeded in differentiating hepatocytes from ES cells without EB formation, using simple adherent monoculturing in media comprising several growth factors in culture dishes with two different matrices.
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Citations
21 Claims
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1. A method for inducing differentiation of pluripotent cells comprising the following steps (a) and (b):
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(a) culturing the pluripotent cells in a medium comprising any one of the following growth factors (i) to (iii);
(i) acidic fibroblast growth factor, fibroblast growth factor 4, and hepatocyte growth factor;
(ii) acidic fibroblast growth factor, and growth factor(s) selected from activin A, epidermal growth factor, and β
-nerve growth factor; and
(iii) fibroblast growth factor 4, and growth factor(s) selected from activin A and hepatocyte growth factor; and
,(b) culturing the cell cultured in step (a) in a medium comprising oncostatin M. - View Dependent Claims (2, 3, 5, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18)
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4. A method for inducing differentiation of pluripotent cells comprising the following steps (a) and (b):
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(a) culturing the pluripotent cells in a medium comprising at least one growth factor selected from retinoic acid, leukemia inhibitory factor, and hepatocyte growth factor; and
,(b) culturing the cell cultured in step (a) in a medium comprising any one of the following growth factors (i) to (iii);
(i) acidic fibroblast growth factor, fibroblast growth factor 4, and hepatocyte growth factor;
(ii) acidic fibroblast growth factor, and growth factor(s) selected from activin A, epidermal growth factor and β
-nerve growth factor; and
(iii) fibroblast growth factor 4, and growth factor(s) selected from activin A and hepatocyte growth factor.
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6. A method for inducing differentiation of pluripotent cells comprising the following steps (a) to (c):
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(a) culturing the pluripotent cells in a medium comprising at least one of the growth factors selected from retinoic acid, leukemia inhibitory factor and hepatocyte growth factor;
(b) culturing the cell cultured in step (a) in a medium comprising any one of the following growth factors (i) to (iii);
(i) acidic fibroblast growth factor, fibroblast growth factor 4 and hepatocyte growth factor;
(ii) acidic fibroblast growth factor, and growth factor(s) selected from activin A, epidermal growth factor and α
-nerve growth factor; and
(iii) fibroblast growth factor 4, and growth factor(s) selected from activin A and hepatocyte growth factor; and
,(c) culturing the cells cultured in step (b) in a medium comprising oncostatin M. - View Dependent Claims (7)
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19. A kit comprising any one of the following (a) to (c):
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(a) acidic fibroblast growth factor, fibroblast growth factor 4, and hepatocyte growth factor;
(b) acidic fibroblast growth factor, and growth factor(s) selected from activin A, epidermal growth factor, and β
-nerve growth factor; and
(c) fibroblast growth factor 4, and growth factor(s) selected from activin A and hepatocyte growth factor. - View Dependent Claims (20, 21)
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Specification