Human hepatocyte-like cells and uses thereof
First Claim
1. A method for preparing human hepatocyte-like cells, which comprises the steps of:
- (a) incubating undifferentiated cells derived from human in a culture medium containing a combination of growth factors selected from (i) to (iii) below;
(i) acidic fibroblast growth factor, fibroblast growth factor 4, and hepatocyte growth factor;
(ii) acidic fibroblast growth factor and a growth factor selected from the group consisting of activin A, epidermal growth factor, and β
-nerve growth factor; and
(iii) fibroblast growth factor 4 and a growth factor selected from the group consisting of activin A and hepatocyte growth factor;
;
(b) incubating the cells cultured in step (a) in a culture medium containing oncostatin M; and
(c) incubating the cells cultured in step (b) in a culture medium containing dexamethasone, wherein the total culturing period is about 2 to about 13 weeks.
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Abstract
The present invention provides modified methods of producing human hepatocyte-like cells which exhibit phenotypes more similar to those of human hepatocytes. The present invention also provides the human hepatocyte-like cells and uses thereof. The methods of the present invention comprises a prolonged incubation period and addition of dexamethasone to the culture medium. The methods of the inventors produce human hepatocyte-like cells whose morphology is more similar to that of human hepatocytes, compared with the conventional system. The cells produced have both morphological and functional features of primary culture cells from normal human liver, including cytochrome P450 (CYP), multi-drug resistance-associated protein (MRP), and multi-drug protein (MDR). The use of human hepatocyte-like cells of the present invention enables, without using any animal model, assessment of metabolism and hepatotoxicity of test compounds, which are drug candidates, and screening for therapeutic agents for hepatic diseases, inhibitors to hepatitis virus infection, and therapeutic agents for viral hepatitis.
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10 Claims
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1. A method for preparing human hepatocyte-like cells, which comprises the steps of:
-
(a) incubating undifferentiated cells derived from human in a culture medium containing a combination of growth factors selected from (i) to (iii) below;
(i) acidic fibroblast growth factor, fibroblast growth factor 4, and hepatocyte growth factor;
(ii) acidic fibroblast growth factor and a growth factor selected from the group consisting of activin A, epidermal growth factor, and β
-nerve growth factor; and
(iii) fibroblast growth factor 4 and a growth factor selected from the group consisting of activin A and hepatocyte growth factor;
;
(b) incubating the cells cultured in step (a) in a culture medium containing oncostatin M; and
(c) incubating the cells cultured in step (b) in a culture medium containing dexamethasone, wherein the total culturing period is about 2 to about 13 weeks. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10)
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Specification