Extended Data Fig. 6: HepOrg in assembloid media form narrow and homogenous bile canaliculi network.
From: Mouse liver assembloids model periportal architecture and biliary fibrosis
a. Immunofluorescence staining for bile canaliculi (CD13, green), nuclei (DAPI, blue) and cell borders (Phalloidin, grey) in HepOrg grown in the different media. Representative images from 3 independent experiments are shown, Scale bar, 100 µm. b. Immunofluorescence staining for bile canaliculi (CD13, green) and cell borders (Phalloidin, grey) in liver tissue (left), HepOrg cultures in MM for 7 days (middle), or HepOrg grown in HM-Wnt media (right). Representative images are shown of n = 3 independent experiments. Scale bar, 20 µm, zoom-in, 10 µm. c. Representative BC networks from healthy mouse liver tissue (left), HepOrg grown in MM media (middle), and HepOrg in HM-Wnt media (right). Colour corresponds to the mean bile canaliculi (BC) diameter in μm as indicated in intensity scale (blue, BC < 1.5 um; white, BC > 6 um). Note that BC is most homogenous in tissue, followed by HepOrg in MM media, while HepOrg in HM-Wnt media show large BC diameter variability. n = 3 independent experiments. d. Still images from time-lapse imaging analysis of fluorescent phosphatidylcholine (16:0-06:0 NBD PC) confirms functionality of MDR2 transporter. Compounds are shown in Royal LUT. Nuclei (SPY555-DNA, magenta) and actin (SiR-act, cyan) are also shown. n = 3 independent experiments. Scale bar, 50 µm. e. Dot plot shows gene expression from scRNAseq of HM-Wnt and MM media HepOrg for bile transporter genes.
