In Vitro Generation of Hepatocytes from Human Embryonic Stem Cells
First Claim
1. A differentiated cell population in an in vitro culture generated by differentiating human pluripotent stem cells in HepG2 conditioned medium.
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Abstract
Differentiation of human pluripotent stem cells, such as human embryonic stem cells (hESC), into hepatocytes by in vitro methods is disclosed. The pluripotent stem cells are cultured in conditioned medium from the hepatocarcinoma cell line, HepG2. Specific growth factors and defined media may also be added to the medium for stage specific differentiation of the derived hepatocytes. Hepatocytes differentiated from human pluripotent stem cells may be characterized by fluorescence activated cell sorting (FACS), immunofluorescence analysis (IF), real time polymerase reaction (RT-PCR), and functional assays. The methods disclosed herein are able to differentiate high percentages of hepatocytes from human pluripotent stem cells using the disclosed methods. These differentiated cells may exhibit polygonal shape morphology, typical of hepatocytes, and may express hepatocyte specific genes. The differentiated cells may also be positive for definitive endoderm markers and hepatic markers.
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21 Claims
- 1. A differentiated cell population in an in vitro culture generated by differentiating human pluripotent stem cells in HepG2 conditioned medium.
- 8. A method of generating a differentiated cell population from human pluripotent stem cells comprising maintaining a culture of undifferentiated human pluripotent stem cells and culturing the culture of undifferentiated human pluripotent stem cells in the presence of HepG2 conditioned medium.
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14. A method of generating a differentiated cell population from human embryonic stem cells comprising:
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a) maintaining a culture of undifferentiated human embryonic stem cells; b) culturing the undifferentiated human embryonic stem cells in the presence of HepG2 conditioned medium; c) maintaining the culture in a basal medium; and d) maintaining the culture in hepatocyte culture media; wherein the differentiated cell population generated expresses at least one hepatocyte marker. - View Dependent Claims (15)
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