Fig. 4: IL6 derived iMHs repopulate livers of Fah^−/− mice.

a Schematic diagram of the experimental procedure: td-iMHs matured from IL6-expanded td-iHPCs (mixed population or single-cell clones) were transplanted into the liver of Fah^−/− mice, and the repopulated td-iMHs were isolated again 8 weeks later and tested for 2nd round of expansion with IL6. b Body weight change of Fah^−/− mice (no NTBC) receiving vehicle (Non-trans, n = 9 mice), primary td-hepatocytes (td-Hepa, n = 11 mice), IL6-td-iMHs (P10, n = 16 mice) and IL6-td-iMHs-clone-1 (P10, n = 18 mice). Data are Means ± SEM. Statistical differences between two groups were analyzed from D0–D28. ***p < 0.001 (two-way ANOVA). c Survival curves of the animals in b. ***p < 0.001 (Log-rank test). d Representative images of the whole livers (top), and fluorescent images of the whole livers (middle, scale bars represent 5 mm) and frozen sections of liver (down, scale bars represent 1 mm) from Fah^−/− mice before NTBC withdrawal (D0, n = 2 mice), 28 days after vehicle transplantation (no NTBC, n = 3 mice), or 63 days after td-Hepa (n = 3 mice), IL6-td-iMHs (n = 6 mice), or IL6-td-iMHs-clone-1 (n = 7 mice) transplantation (no NTBC). The liver of Alb-td-mice was used as control. e Quantitative analysis of tdTomato positive areas in liver (n = 2–7 mice, numbers of the animals were listed on top of the bar). f Serum levels of ALT, AST, ALP and TBIL in the Fah^−/− mice after transplantation. Data are Means ± SEM (n = 2–17 mice, numbers of the animals were listed under the bar). **p < 0.01, ***p < 0.001 (two-tailed unpaired Student’s t test). g Immunofluorescence staining of hepatic marker Fah (green) in frozen liver sections of Fah^−/− mice before or 63 days after transplantation. Nuclei were stained with Hoechst 33342 (blue). Representative morphology and tdTomato images (h) and growth curves (i) of hepatocytes isolated from the Fah^−/− mice repopulated with IL6-td-iMHs for 63 days and then cultured in vitro with or without IL6 for 14 days. Data are Means ± SEM (n = 4 independent experiments). ***p < 0.001 (two-way ANOVA). Scale bars represent 100 µm. ALT alanine aminotransferase, AST aspartate aminotransferase, ALP alkaline phosphatase, TBIL total bilirubin. Source data and exact p values are provided in a Source data file.