Human Pluripotent Stem Cell-Derived Organoids as Models of Liver Disease

Bin Ramli MN, Lim YS, Koe CT, Demircioglu D, Tng W, Uy Gonzales KA, Tan CP, Szczerbinska I, Liang H, Soe EL, Lu Z, Ariyachet C, Yu KM, Koh SH, Yaw LP, Binte Jumat NH, Yew Lim JS, Wright G, Shabbir A, Dan YY, Ng H-H, Chan Y-S
Source: World J Gastroenterol
Publication Date: (2020)
Issue: 159(4): 1471-1486
Research Area:
Cells used in publication:
Induced Pluripotent Stem Cell (iPS), human
Species: human
Tissue Origin:
Culture Media:

Visualization of functional bile canaliculi

All organoids utilized for bile canaliculi visualization were subjected to standard preparation consisting of a single wash with DPBS containing calcium and magnesium (DPBS Ca/Mg) (Gibco) followed by Hoescht staining in Clonetics™ HCM™ Hepatocyte Culture Medium (HCM) (Lonza) for 30 mins at 37°C. For snapshot imaging of the bile canaliculi, organoids were incubated in HCM media containing 1uM of CDFDA (Molecular Probes) for 2h at 37°C, washed twice with DPBS Ca/Mg and transferred into a well of a µ-slide Angiogenesis glass bottom plate (Ibidi) containing HCM. For time-lapse imaging, organoids were kept constantly in HCM media containing 1uM of CDFDA (HCM-CDFDA). Organoids were incubated with indicated concentrations of troglitazone (Sigma Aldrich) at both 37°C and room temperature as specified.


We cultured H1 human embryonic stem cells (WA-01, passage 27-40) and induced pluripotent stem cells (GM23338) with a series of chemically defined and serum-free media to induce formation of posterior foregut cells, which were differentiated in 3-dimensions into hepatic endoderm spheroids and stepwise into hepatoblast spheroids. Hepatoblast spheroids were re-seeded in a high-throughput format and induced to form hepatic organoids; development of functional bile canaliculi was imaged live. We also compared gene expression profiles between liver tissue samples from patients with nonalcoholic steatohepatitis (NASH) vs without. We quantified hepatocyte and cholangiocyte populations in organoids using immunostaining and flow cytometry; cholangiocyte proliferation of cholangiocytes was measured. We compared the bile canaliculi network in the organoids incubated with vs without free fatty acids by live imaging.