In vitro hepatic maturation of human embryonic stem cells by using a mesenchymal cell line derived from murine fetal livers
- 244 Downloads
Hepatocytes derived from human embryonic stem cells (hESCs) are an attractive cell source for regenerative medicine. We previously reported the differentiation of hESCs into alpha-fetoprotein (AFP)-producing endodermal cells by using extracellular matrix and growth factors. We also reported the establishment of the MLSgt20 cell line, which was derived from mesenchymal cells residing in murine fetal livers and accelerated the hepatic maturation of both murine hepatic progenitor cells and murine ESCs. In this study, hESC-derived AFP-producing cells were isolated by using a flow cytometer and co-cultured with MLSgt20 cells. The co-cultured hESC-derived AFP-producing cells had the immunocytological characteristics of hepatocytes, expressed mature hepatocyte markers (as indicated by reverse transcription and the polymerase chain reaction), and displayed higher hepatocyte functions including ammonia removal, cytochrome P450 3A4/7 activity, and the ability to produce and store glycogen. However, the MLSgt20 cells did not directly cause undifferentiated hESCs to mature into hepatocyte-like cells. The co-culture method was thus successfully shown to induce the differentiation of hESC-derived endodermal cells into functional hepatocyte-like cells.
KeywordsEmbryonic stem cells Alpha-fetoprotein Hepatocyte MLSgt20 cell line CYP3A Mouse
We thank Dr. Andrew G. Farr for providing the anti-gp38 antibodies.
- Ek M, Soderdahl T, Kuppers-Munther B, Edsbagge J, Andersson TB, Bjorquist P, Cotgreave I, Jernstrom B, Ingelman-Sundberg M, Johansson I (2007) Expression of drug metabolizing enzymes in hepatocyte-like cells derived from human embryonic stem cells. Biochem Pharmacol 74:496–503CrossRefPubMedGoogle Scholar
- Fukumitsu K, Ishii T, Yasuchika K, Amagai Y, Saitoh M, Kawamoto T, Kawase E, Suemori H, Nakatsuji N, Ikai I, Uemoto S (2009) Establishment of a cell line derived from mouse fetal liver that have the character to promote the hepatic maturation of mouse embryonic stem cells by co-culture method. Tissue Eng Part A (in press)Google Scholar
- Gellner K, Eiselt R, Hustert E, Arnold H, Koch I, Haberl M, Deglmann C, Burk O, Buntefuss D, Escher S, Bishop C, Koebe H, Brinkmann U, Klenk H, Kleine K, Meyer U, Wojnowski L (2001) Genomic organization of the human CYP3A locus: identification of a new, inducible CYP3A gene. Pharmacogenetics 11:111–121CrossRefPubMedGoogle Scholar
- Kamo N, Yasuchika K, Fujii H, Hoppo T, Machimoto T, Ishii T, Fujita N, Tsuruo T, Yamashita JK, Kubo H, Ikai I (2007) Two populations of Thy1-positive mesenchymal cells regulate in vitro maturation of hepatic progenitor cells. Am J Physiol Gastrointest Liver Physiol 292:G526–G534CrossRefPubMedGoogle Scholar