Patent ID: 9057051
Filing Date: 2015-06-16
Classification: C12N

Abstract:
1. A method for inducing cells to differentiate into cells that express a hepatocyte phenotype, said method comprising the steps of: (a) culturing either mouse or human embryonic stem (ES) cells, mouse or human induced pluripotent stem (iPS) cells or mouse, rat, or human multipotent adult progenitor cells characterized in that the multipotent adult progenitor cells can differentiate into at least one cell type of each of the endodermal, ectodermal, and mesodermal embryonic lineages but are not embryonic germ cells, embryonic stem cells, or germ cells, with about 5 ng/ml to about 500 ng/ml Wnt3a and about 10 ng/ml to about 1,000 ng/ml ActivinA, to obtain cells that express a definitive endoderm phenotype, (b) then culturing the cells produced in step (a) with about 1 ng/ml to about 100 ng/ml bFGF and about 5 ng/ml to about 5 ng/ml to about 50 ng/ml BMP4, to obtain cells that express a liver committed endodermal phenotype, (c) then culturing the cells produced in step (b) with about 5 ng/ml to about 500 ng/ml aFGF, about 1 ng/ml to about 100 ng/ml FGF4 and about 2.5 ng/ml to about 250 ng/ml FGF8b, to obtain cells that express a hepatoblast phenotype, and (d) then culturing the cells produced in step (c) with about 2 ng/ml to about 200 ng/ml HGF and about 10 ng/ml to about 1,000 ng/ml Follistatin, to obtain cells that express a hepatocyte phenotype.