Fig. 5

Lgr5⁺ liver stem cells existed in human fibrotic livers. a Lgr5 staining in human liver samples using anti-Lgr5 antibody. b Lgr5 mRNA level in human liver samples. c FACS plot for human Lgr5⁺ liver cells (left) and a schematic overview of the experimental setup (top right). Single Lgr5⁺ cells were isolated from six independent donors human liver biopsies diagnosed with liver fibrosis, they were cultured for organoids formation, and continuously cultured for at least three passages in vitro. The representive images were shown (bottom right). d–f Single Lgr5⁺ cells were isolated from six independent donors human liver biopsies diagnosed with liver fibrosis and cultured, and different concentrations of rHGF/rRspo1 were added into the culture medium; the representative organoid pictures, organoids number, and organoids size were analyzed (d). The Lgr5 mRNA level in the organoids was measured using qPCR assay (e). The Lgr5⁺ cells numbers were measured using FACS assay (f). For a, e, triplicates for each condition were analyzed. The results are shown as mean ± s.d. of three independent experiments. *p < 0.05. For d, f, the results are shown as mean ± s.d. of three independent experiments. *p < 0.05. Scale bars, 200 μm (a), 250 μm (c, d)