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Culturing conditions conducive to the differentiation of hESCs along a hepatic lineage
Author(s) -
Zamule Stephanie Mary,
Weyant Denise M,
Omiecinski Curtis J
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.22.1_supplement.921.11
Subject(s) - embryonic stem cell , hepatocyte , biology , microbiology and biotechnology , andrology , biochemistry , gene , medicine , in vitro
The limited availability of hepatic tissue suitable for the treatment of liver disease and for research makes the generation of hepatic‐like cells from alternative sources an attractive proposition. Although a number of studies have demonstrated that human embryonic stem cells (hESCs) are capable of differentiating into hepatic precursors, the methodology has yet to be fully elucidated. To this end, we cultured hESCs for 10 days on collagen substrata in our primary hepatocyte maintenance media comprised of William's E media supplemented with 25 nM dexamethasone, 10 nM insulin, 5 ng/ml selenium, 5 μg/ml transferrin, and 1% linoleic acid. Using quantitative RT‐PCR assays, we found that hESCs cultured under these conditions exhibited a 2‐fold decrease in mRNA levels of the stem cell pluripotency marker, Oct4, a ~30,000‐fold increase in levels of the endodermal marker, α‐fetoprotein, and a ~475‐fold increase in the hepatic marker, albumin. Further, levels of the constitutive androstane receptor, CAR, were increased ~530‐fold, to levels comparable to those in primary cultures of human hepatocytes. These results define a novel, straightforward methodology that may be useful in facilitating hESC differentiation along the hepatic lineage within a relatively short period of time and provide a framework for subsequent studies seeking to develop a renewable supply of human hepatocytes for research applications. Supported by NIH grant, GM66411.