Functional hepatocyte-like cells derived from mouse embryonic stem cells: a novel in vitro hepatotoxicity model for drug screening.
The aim of the present study was to differentiate mouse embryonic stem (mES) cells into a high percentage of hepatocyte-like cells, and to demonstrate their utility as an in vitro hepatotoxicity model. We were able to differentiate 80-90% of mES cells using optimized hepatocyte differentiation medium. These differentiated cells showed typical hepatocyte morphology, expressed hepatic specific genes as shown by RT-PCR and displayed antibody detectable expression of markers specific for hepatic maturation. These hepatocyte-like cells also demonstrated evidence of glycogen storage. These cells when exposed to CCl4, a commonly used hepatotoxicant, showed an elevation of liver function enzymes, SGOT, SGPT and LDH, indicating hepatic damage. Further, this increase was prevented by pre-treatment with N-acetylcysteine, a known anti-oxidant. Thus we propose that the hepatocyte-like cells derived by the present method may prove to be useful as an in vitro model of hepatotoxicity, thereby providing a novel and promising alternative for obtaining large numbers of functional hepatocyte-like cells for in vitro drug metabolism and hepatotoxicity screening of potential drug candidates.
Kulkarni
J S
JS
Embryonic Stem Cell Group, Reliance Life Sciences Pvt Ltd., Dhirubhai Ambani Life Sciences Centre, R-282, TTC Industrial Area of MIDC, Thane Belapur Road, Rabale, Navi Mumbai 400 071, India.
Khanna
A
A
D000410:Alanine Transaminase / Q000378:metabolism; D000818:Animals; D001219:Aspartate Aminotransferases / Q000378:metabolism; D002251:Carbon Tetrachloride / Q000633:toxicity; D002454:Cell Differentiation; D002478:Cells, Cultured; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D004622:Embryo, Mammalian / Q000166:cytology*; D005455:Fluorescent Antibody Technique; D022781:Hepatocytes / Q000187:drug effects*; D006801:Humans; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D051379:Mice; D012333:RNA, Messenger / Q000032:analysis; D013234:Stem Cells / Q000166:cytology*
D012333:RNA, Messenger; D002251:Carbon Tetrachloride; D007770:L-Lactate Dehydrogenase; D001219:Aspartate Aminotransferases; D000410:Alanine Transaminase