Method of deriving mature hepatocytes from human embryonic stem cells
AssigneeWisconsin Alumni Research Foundation
InventorsSrikumar Sengupta, James Thomson
Life of this patent
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Abstract
A method for producing mature hepatocytes having functional hepatic enzyme activity from human pluripotent cells is disclosed. The method includes the step of transferring an external vector that includes a microRNA having the seed sequence of the microRNA miR-122, a DNA sequence coding for such a microRNA, a microRNA having the seed sequence of the microRNA miR-let-7c, a DNA sequence coding for such a microRNA, or a combination these, into one or more fetal hepatocytes. The resulting cells differentiate into mature hepatocytes that exhibit functional hepatic enzyme activity, and that can be used in drug metabolism and toxicity testing, in the study of viruses that target hepatic tissue, and as therapeutics.
Claims12
A method for producing mature hepatocytes from human pluripotent stem cells, the method comprising the step of transferring into one or more fetal hepatocytes obtained from human pluripotent cells:
- (a) a first microRNA comprising a core sequence that is 18 to 24 nucleotides long, wherein the 10 nucleotide sequence on the 5′ end of the core sequence comprises a seed sequence consisting of any 6, 7, 8, 9 or 10 consecutive nucleotides of SEQ ID NO:5;
- or
- (b) an external vector comprising a DNA sequence coding for the first microRNA;
- or
- (c) a second microRNA comprising a core sequence that is 18 to 24 nucleotides long;
- wherein the 10 nucleotide sequence on the 5′ end of the core sequence comprises a seed sequence consisting of any 6, 7, 8, 9 or 10 consecutive nucleotides of SEQ ID NO:6;
- or
- (d) an external vector comprising a DNA sequence coding for the second microRNA;
- or
- (e) two or more of (a)-(d);
- wherein (c) the second microRNA, (d) the external vector comprising a DNA sequence coding for the second microRNA, or both (c) and (d) are transferred into the one or more fetal hepatocytes, and whereby the cells differentiate into mature hepatocytes exhibiting functional hepatic enzyme activity.
Dependencies are inferred from the claim text.