efficient differentiation of embryonic stem cells into hepatic cells in vitro using a feeder-free basement membrane substratum高效的胚胎干细胞分化成肝细胞在体外使用feeder-free基底膜下层.pdfVIP

Efficient Differentiation of Embryonic Stem Cells into Hepatic Cells In Vitro Using a Feeder-Free Basement Membrane Substratum 1,2 1,2 3 3 3 Nobuaki Shiraki , Taiji Yamazoe , Zeng Qin , Keiko Ohgomori , Katsumi Mochitate , Kazuhiko Kume1,2, Shoen Kume1,2* 1 Department of Stem Cell Biology, Institute of Molecular Embryology and Genetics, Kumamoto University, Honjo, Kumamoto, Japan, 2 Global Center of Excellence Program, Kumamoto University, Honjo, Kumamoto, Japan, 3 BM Matrix Laboratory, Environmental Health Sciences Division, National Institute for Environmental Studies, Ibaraki, Japan Abstract The endoderm-inducing effect of the mesoderm-derived supportive cell line M15 on embryonic stem (ES) cells is partly mediated through the extracellular matrix, of which laminin a5 is a crucial component. Mouse ES or induced pluripotent stem cells cultured on a synthesized basement membrane (sBM) substratum, using an HEK293 cell line (rLN10-293 cell) stably expressing laminin-511, could differentiate into definitive endoderm and subsequently into pancreatic lineages. In this study, we investigated the differentiation on sBM of mouse and human ES cells into hepatic lineages. The results indicated that the BM components played an important role in supporting the regional-specific differentiation of ES cells into hepatic endoderm. We show here that knockdown of integrin b1 (Itgb1) in ES cells reduced their differentiation into hepatic lineages and that this is mediated through Akt signaling activation. Moreover, under optimal conditions, human ES cells differentiated to express mature hepatocyte markers and secreted high levels of albumin. This novel procedure for inducing hepatic differ