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US 12,734,197 B2Granted
Generation of uniform hepatocytes from human embryonic stem cells by inhibiting TGF-beta and methods of maintaining hepatic cultures
AssigneeWisconsin Alumni Research Foundation
InventorsJames A. Thomson, Srikumar Sengupta
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Abstract
This disclosure relates generally to new methods of maintaining the expression of hepatic genes in human hepatocytes and method for maintaining the functional hepatic enzyme activity of primary hepatocytes in culture. The disclosure also encompasses new methods of deriving a population of pure hepatocytes without selecting or sorting the cells from the cultured pluripotent cells.
Claims11
A method of expressing or inducing expression of one or more mature hepatic gene(s) or maintaining expression thereof in a human hepatocyte, the method comprising:
- culturing the human hepatocyte with at least one inhibitor of mitogen activating protein kinase (MAPK),
- wherein the one or more mature hepatic gene(s) are Bile Acid-CoA:Amino Acid N-Acyltransferase (BAAT), Solute Carrier Organic Anion Transporter Family Member 1B1 (SLCO1B1), Cytochrome P450 Family 1 Subfamily B Member 1 (CYP1B1), Cytochrome P450 Family 2 Subfamily C Member 18 (CYP2C18), UDP Glucuronosyltransferase Family 1 Member A6 (UGTIA6), UDP-glucuronosyltransferase 1-9 (UGT1A9), and Cluster of Differentiation 81 (CD81);
- and
- wherein the cultured human hepatocyte expresses the one or more mature hepatic gene(s) for at least 72 hours at an expression level that is at least 70% the expression level of the same gene(s) in fresh primary hepatocytes.
1 more element
Dependencies are inferred from the claim text.