Hepatocyte-like cells

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. 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.