Recombinant human oncostatin m

Following established protocols (Cai et al., 2008 ), control, LMNA T10I, LMNA R541C hiPSCs were grown in feeder-free differentiation conditions. For efficient hepatocyte differentiation, cells were incubated in definitive endoderm media with recommended supplements (Stem Cell Technologies) for 4 days in ambient O₂ and 5% CO₂, yielding homogeneous monolayer of definitive endoderm cells. At day 5, cells were incubated with recombinant human BMP-4 (20ng/mL; Peprotech) and recombinant human FGF basic (10ng/mL; R&D Systems) for 5 days in RPMI-B27 (with insulin) in 5% O₂ and 5% CO₂, yielding hepatic progenitor cells. At day 10, cells were incubated in RPMI-B-27 (with insulin) supplemented with recombinant human HGF (20ng/mL; PeproTech) for 5 days at 5% O₂ and 5% CO₂, yielding immature HLCs. Finally, at d15, cells were incubated with HCM Hepatocyte Culture Medium (Lonza) without EGF and supplemented with recombinant human oncostatin M (20ng/mL; R&D Systems) for 7 days in ambient O₂ and 5% CO₂, yielding mature HLCs, which were collected at day 23 for subsequent ChIP, immunofluorescence, and immunoblotting.