Promoted differentiation of cynomolgus monkey ES cells into hepatocyte-like cells by co-culture with mouse fetal liver-derived cells

Figure 1 Protocol for differentiation of cES cells. A: Spontaneous differentiation. Undifferentiated cES cells were cultured in gelatin-coated dishes in DMEM basic medium for 28 d without additional feeders or growth factors; B: GF-induced differentiation. Undifferentiated cES cells were cultured in DMEM basic medium containing LIF (100 units/mL) and RA (10^-8 mol/L) for 3 d, followed by a 5-d culture in DMEM basic medium containing FGF2 (10 mg/L), FGF4 (20 mg/L), HGF (25 mg/L), and OSM (10 mg/L). Thereafter, the cells were cultured for 20 d in DMEM basic medium containing 10^-7 mol/L dexamethasone (Dex), 0.2 mmol/L, L-ascorbic-2-phosphate (AP), and 10 mmol/L nicotinamide (NA); C: Promoted differentiation by co-culture with MFLCs. Undifferentiated cES cells were plated onto 60-mm dishes and co-cultured with MFLCs in a 35-mm culture insert across a 0.4-μm Millicell CM membrane for 28 d.