Figure 7.

Generation of human 3-dimensional (3-D) organoids and assessment of apical sodium bile acid (BA) transporter (ASBT)/chymase/histamine/fibroblast growth factor (FGF) signaling. Control and primary sclerosing cholangitis (PSC) cholangiocytes (from n = 1 patient per group) were isolated, immortalized, and used for organoid formation as detailed in the schematic created with bioRender.com (A). Human PSC organoids have enhanced ASBT (red) immunoreactivity, costained with cytokeratin-19 (CK-19, green) and desmin (yellow) to mark cholangiocytes and hepatic stellate cells (HSCs), respectively, compared with control organoids (B). Furthermore, histamine (C) and FGF19 (D) secretion were measured in PSC organoids compared with human control organoids. Chymase (red) immunoreactivity (E) was detected in control and PSC organoids. Data are presented as mean ± SD of 4 and 5 experiments for histamine and FGF19 enzyme immunoassay (EIA), respectively, from 25 organoids created from 1 human PSC patient and 1 human control patient. Representative images are presented at ×10 with ×40 zoom from 1 organoid per group.