Fig.3

SHED-HepT reconstructs intrahepatic bile ducts in livers of recipient CCl₄-treated mice. a–e In vitro hepatobiliary function was analyzed in SHED-Heps. Gene expression of uridine 5′-diphospho-glucuronosyltransferase 1A1 (UGT1A1) by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Amount of direct bilirubin by colorimetric assay (b). Representative images of bile transport were detected by CLF staining. Nuclei were stained with DAPI (c). Gene expression of membrane metalloendopeptidase (MME), ATP-binding cassette transporter B1 (ABCB1), ABCB11, and ABCC2 by RT-qPCR d. Representative images of the expression of MME, ABCB1, ABCB11, and ABCC2 were detected by immunohistochemical analysis (e). a–e hPHep, human primary hepatocyte. a, b, d n = 5 for all groups. ***P < 0.005. nd, no detection. The graph bars represent the means ± SEM. a, d Results are shown as a ratio to hPHeps (hPHep = 1). c, e Scale bars, 20 μm. f, g Representative histograms of the expression of cell surface markers for hepatocyte progenitors and mesenchymal stem cells in SHED-Heps (f) and whole liver cells isolated from recipient mice (WLC) (g) were detected by FCM assay. EPCAM: epithelial cell adhesion molecule; MME: membrane metalloendopeptidase; NCAM1: neural cell adhesion molecule; PROM1: promin 1; R-PE: R-phycoerythrin. f Red solid line, target antibody-stained histograms; black solid line, isotype-matched control-stained histograms. g Area filed with red, target antibody-stained histograms; solid line, isotype-matched control-stained histograms