Differentiation of hiPSCs into Mature Hepatocytes

Human‐inducible pluripotent stem cells (hiPSCs) were cultured with mTESR1 medium in a Matrigel‐coated culture dish. After the cell density reached 70%, the hİPSC medium was replaced with Roswell Park Memorial Institute / B27 medium containing 100 ng/mL Activin A (PeproTech), 50 ng/mL Wnt3a(R&D Systems), and 10 ng/mL HGF(PeproTech) for 3–5 days to induce endoderm cells. Subsequently, endodermal cells were differentiated to progenitor cells by changing RPMI medium with the knockout [KO]/DMEM containing 20% knockout serum replacement, 1 mml‐glutamine, 1% nonessential amino acid, 0.1 mm 2‐mercaptoethanol, and 1% dimethyl sulfoxide for further 4 days. In the final stage, cells were cultured with Iscove’s modified Dulbecco's (IMDM) medium containing 20 ng/mL oncostatin M (Invitrogen), 0.5 lM dexamethasone, and 50 mg/mL ITS premix(BD Biosciences) for 5 days in order to obtain mature hepatocytes [22 (link), 23 (link)]