Generating hepatocytes

摘要

Compositions and methods are described herein for inducing reprogramming of non-pluripotent cells across lineage and differentiation boundaries to generate endodermal progenitor cells and hepatocytes. Compositions and methods for expansion of endodermal progenitor cells without loss of phenotype are also described herein.

主权项

1. A method comprising administering to a subject one or more immature hepatocytes generated as follows:
(a) incubating one or more cells with a first composition comprising an epidermal growth factor (EGF), a basic fibroblast growth factor (bFGF), an activator of WNT signaling, a nuclear receptor liver receptor homolog 1 agonist, a histone deacetylase (HDAC) inhibitor, a histone demethylase LSD1 inhibitor, and a DNA methyltransferase (DNMT) inhibitor to generate induced multipotent cells; (b) incubating the induced multipotent cells with a second composition comprising an activator of WNT signaling, a nuclear receptor liver receptor homolog 1 agonist, a histone deacetylase (HDAC) inhibitor, a histone demethylase LSD1 inhibitor, a DNA methyltransferase (DNMT) inhibitor, Activin A, or any combination thereof to generate one or more endodermal progenitor cells; (c) optionally incubating the one or more endodermal progenitor cells in a third composition comprising epidermal growth factor (EGF), basic fibroblast growth factor (bFGF), and a TGF-beta inhibitor for a time and in a sufficient amount of the third composition to generate an endodermal progenitor cell population; and (d) incubating the one or more endodermal progenitor cells or the endodermal progenitor cell population in a fourth composition comprising basic fibroblast growth factor (bFGF), a TGF-beta inhibitor, bone morphogenetic protein 4 (BMP4), and dexamethasone for a time and in a sufficient amount of the composition to generate one or more immature hepatocytes.