Enrichment of pluripotent stem cell-derived hepatocyte-like cells

The gene expression profiles of differentiating human pluripotent stem cells (hPSCs) were analyzed by DNA microarray. We identified differences and commonalities among six human pluripotent stem cell lines: the hESCs KhES1, KhES2, KhES3, and H1, and the iPSCs 201B7 and 243G1. Analysis of DNA microarray data suggested that hepatocyte-like differentiation of EBs treated with ammonia in Lanford medium. Undifferentiated pluripotent stem cells were prepared using the standard method with MEF-coated dishes (ML) or MEF-conditioned medium method with matrigel-coated dishes (MC). EB were formed by transferring small clumps (200 µm diameter) of hPSCs to a non-adhesive 60-mm dish coated with Lipidure. Suspension cultures were maintained for 5 days. Following EB formation, 1 µL/mL of 10% w/v aqueous ammonia (Wako) was added to fresh Lanford medium. This process was repeated every 2 days for 14 days total.