RRC ID 27180
Author Meng Q, Haque A, Hexig B, Akaike T.
Title The differentiation and isolation of mouse embryonic stem cells toward hepatocytes using galactose-carrying substrata.
Journal Biomaterials
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.
Volume 33(5)
Pages 1414-27
Published 2012-2-1
DOI 10.1016/j.biomaterials.2011.11.007
PII S0142-9612(11)01345-7
PMID 22118818
MeSH Adsorption / drug effects Animals Asialoglycoprotein Receptor / metabolism Cadherins / metabolism Cell Differentiation / drug effects* Cell Separation / methods* Cells, Cultured Disaccharides / pharmacology* Embryonic Stem Cells / cytology* Embryonic Stem Cells / drug effects* Embryonic Stem Cells / metabolism Endoderm / cytology Endoderm / drug effects Endoderm / metabolism Galactose / pharmacology* Hepatocytes / cytology* Hepatocytes / drug effects Hepatocytes / metabolism Immobilized Proteins / metabolism Mice Phenotype Polystyrenes / pharmacology Receptors, Fc / metabolism Surface Properties / drug effects Vinyl Compounds / pharmacology*
IF 10.273
Times Cited 25
DNA material Mouse E-cadherin full length cDNA (RDB01184)