title: The differentiation and isolation of mouse embryonic stem cells toward hepatocytes using galactose-carrying substrata.
abstract: A simple culture system to achieve the differentiation of embryonic stem (ES) cells toward hepatocytes with high efficiency is crucial in providing a cell source for the medical application. In this study, we report the effect of a matrix-dependent enrichment of ES cell-derived hepatocytes using immobilized poly(N-p-vinylbenzyl-4-O-β-D-galactopyranosyl-D-gluconamide) (PVLA) with E-cadherin-IgG Fc (E-cad-Fc) as a galactose-carrying substratum. PVLA and E-cad-Fc were confirmed to be stably co-adsorbed onto polystyrene surface by quartz crystal microbalance (QCM). We showed that the E-cad-Fc/PVLA hybrid substratum was efficient in culturing primary hepatocytes and maintaining liver functions, on which the undifferentiated ES cells also maintained high proliferative capability. Furthermore, ES cell-derived hepatocytes on this hybrid matrix expressed elevated level of liver specific genes and functions together with early expression of definitive hepatocyte marker, asialoglycoprotein receptor (ASGPR). Finally, we isolated a high percentage of cells (about 60%) with ASGPR expression after re-seeding onto PVLA-coated surface, and observed the elimination of the poorly differentiated cells (Gata6(+) and Sox17(+)) and the ones toward another cell lineage (brachyury(+) and Pdx1(+)). The system uses a glycopolymer as an extracellular substratum for isolation and enrichment of ES cell-derived hepatocytes with adequate homogeneity and functionality.
authors: Department of Biomolecular Engineering, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Midori-ku, Yokohama, Japan.
authors: Qingyuan
authors: Q
authors: Meng
authors: Amranul
authors: A
authors: Haque
authors: Bayar
authors: B
authors: Hexig
authors: Toshihiro
authors: T
authors: Akaike
mesh_terms: D000327:Adsorption / Q000187:drug effects; D000818:Animals; D037263:Asialoglycoprotein Receptor / Q000378:metabolism; D015820:Cadherins / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects*; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D004187:Disaccharides / Q000494:pharmacology*; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D004707:Endoderm / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D005690:Galactose / Q000494:pharmacology*; D022781:Hepatocytes / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D055767:Immobilized Proteins / Q000378:metabolism; D051379:Mice; D010641:Phenotype; D011137:Polystyrenes / Q000494:pharmacology; D011961:Receptors, Fc / Q000378:metabolism; D013499:Surface Properties / Q000187:drug effects; D014753:Vinyl Compounds / Q000494:pharmacology*
chemical_list: D037263:Asialoglycoprotein Receptor; D015820:Cadherins; D004187:Disaccharides; D055767:Immobilized Proteins; D011137:Polystyrenes; D011961:Receptors, Fc; D014753:Vinyl Compounds; C427524:poly-(N-p-vinylbenzyl-O-galactopyranosyl-(1-4)-gluconamide); D005690:Galactose