Hepatocyte-like cells. Homo sapiens

Hepatocyte-like cells (HLCs) are derived from human pluripotent stem cells (hPSCs) in vitro, but differentiation protocols commonly give rise to a heterogeneous mixture of cells. This variability confounds the evaluation of in vitro functional assays performed using HLCs. We demonstrate the purification of a sub-population of functional HLCs differentiated from multiple hPSC lines using the hepatocyte surface marker Asialoglycoprotein Receptor 1 (ASGR1). We analyzed the expression profile of ASGR1-positive cells by microarray, and tested their ability to perform mature hepatocyte functions (albumin and urea secretion, cytochrome activity). By these measures, ASGR1-positive HLCs are enriched for the gene expression profile and functional characteristics of primary hepatocytes compared to unsorted HLCs. Overall design: We extracted and hybridized RNA to Affymetrix microarrays to obtain gene expression profiles from biological replicates of the following human cell types: hepatocyte-like cells (HLCs) differentiated in vitro from a hESC line and a hiPSC line, FACS-isolated ASGR1+ HLCs differentiated from the same hESC and hiPSC lines, cryopreserved primary human hepatocytes, HepG2 hepatoma cells. HLCs and FACS-isolated ASGR1+ HLCs differentiated from either hESCs or hiPSCs were collected as paired samples.