Figure 1
Functionalization of hepatocyte-like cells derived from hESCs by heat treatment. (A) Schematic representation of the protocol to functionalize hESC-HLCs. d.a.h. represents days after heat treatment. Initially, hPSCs were treated with mTeSR-1 medium supplemented with combinations of Activin A, BMP-4, CHIR99021 (CHIR), LY294002, and Y-27632 to induce definitive endoderm (DE) differentiation until Day 4 (d4). Then, cells were treated with Activin A in RPMI medium supplemented with B27 for anterior definitive endoderm (ADE) specification until Day 8. As the next step, cells were treated with BMP-4 and FGF-10 for inducing hepatocyte commitment in RPMI medium supplemented with B27 until Day 11. Finally, cells were treated with oncostatin M (OSM) and hepatocyte growth factor (HGF) in basal hepatocyte medium for hepatocyte maturation. On day 14, cells were incubated at 37, 39, or 42 °C, and half the total amount of the medium was changed every day. (B) Microscopic images of hESC-HLCs treated with 37, 39, and 42 °C at 6 and 15 d.a.h. Scale bars represent 50 µm. (C) Box plots showing the cell numbers of living hESC-HLCs after heat treatments at 37, 39, and 42 °C at 6 and 15 d.a.h. (n = 4). Centerlines of box plots indicate medians; box limits indicate the 25th and 75th percentiles as determined by R software; whiskers extend 1.5 times the interquartile range from the 25th and 75th percentiles; *p < 0.05, **p < 0.01.
