Figure 3.

CD133+ hBECs transplant in an immunodeficient model of biliary disease

(A) Immunostaining for p53 and p21 in control animals (Oil+DDC) versus induced animals (TM+DDC) (n = 3–4). Scale bars, 120 μm.

(B) Quantification of p53-positive cells per bile duct in tamoxifen (TM), DDC diet (DDC) and induced mice (administered with tamoxifen and DDC diet, TM+DDC). ^∗ denotes p < 0.05 (mean ± SEM), one-way ANOVA (n = 3–4).

(C) Confirmation of cholangiocyte (K19) gene expression in isolated bile ducts (BDs) and absence in hepatocytes (H) normalized to Gapdh. ^∗∗ denotes p < 0.005 (mean ± SEM), Student’s t test (n = 7).

(D) Gene expression analysis of isolated BDs in control (oil) versus induced (TM) mice shows a significant increase in senescent markers in the later population (Trp53, Cdkn1a, Cdkn2a, and Tgfb, normalized to Gapdh). ^∗ denotes p < 0.05, ^∗∗ denotes p < 0.005 (mean ± SEM), Student’s t test (n = 7).

(E) Transaminase analysis in TM, DDC, and induced mice (TM+DDC). ^∗ denotes p < 0.05, ^∗∗∗ denotes p < 0.001, ^∗∗∗∗ denotes p < 0.001 (mean ± SEM), one-way ANOVA (n = 4).

(F) PicroSirius red staining (PiSR) increases in induced mice (TM+DDC), (n = 3). Scale bars, 250 μm.

(G) Representative postmortem image of peritoneum in PBS control and hBEC-transplanted Krt19Cre^ERMdm2^fl/flRag2^−/−Il2rg^−/− mice (n = 4).

(H) Transaminase analysis in PBS control (PBS), human adipose tissue-derived mesenchymal stem cells (hMSCs), and hBEC-transplanted mice (hBEC). ^∗ denotes p < 0.05 (mean ± SEM), one-way ANOVA (n = 5–8).

(I) Survival analysis of mice receiving hBEC (n = 13), PBS (n = 13), or hMSC (n = 5). ^∗ denotes p < 0.05 log-rank (Mantel-Cox) test.

(J) PiSR quantification in PBS control (PBS), hMSC-transplanted mice (hMSC), and hBEC-transplanted mice (hBEC) shows a significant reduction of fibrosis. ^∗ denotes p < 0.05 (mean ± SEM), one-way ANOVA (n = 5).