Fig. 3

Optimizing the HB expanding culture condition. a Schematic of the strategy to identify culture conditions that sustain HB expansion. b Growth curve of HBs cultured in different conditions. Cell growth curves were analyzed by obtaining a cell count. Data are presented as mean ± SEM, n = 3. A: A8301; B₂₀: 20 ng/mL BMP4; B₁₀: 10 ng/mL BMP4; C: CHIR99021; E: EGF; H: HGF; S: SAG. c AFP and HNF4α-positive cells were examined. d Quantitative RT-PCR results showing expression of hepatic genes and Ki67 in expanding HBs. Data are presented as mean ± SEM, n = 3. e Expressions of ALB and E-CAD were measured in the hepatocytes differentiated from HBs expanded with different conditions