Abstract
Cholangiocytes play a crucial role in the pathophysiology of cholestasis. However, research on human cholangiocytes has been restricted by challenges in long-term propagation and large-scale expansion of primary biliary epithelium. The advent of organoid technology has overcome this limitation allowing long-term culture of a variety of epithelia from multiple organs. Here, we describe two methods for growing human cholangiocytes in organoid format. The first applies to the generation of intrahepatic bile ducts using human induced pluripotent stem cells using a protocol of differentiation that recapitulates physiological bile duct development. The second method allows the propagation of primary biliary epithelium from the extrahepatic ducts or gallbladder. Both protocols result in large numbers of cholangiocyte organoids expressing biliary markers and maintaining key cholangiocyte functions.
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Acknowledgments
This work was financially supported by grants by the Addenbrooke’s Charitable Trust (ACT) (F.S.), Academy of Medical Sciences (F.S.), NIHR (F.S.), and the Rosetrees Trust Interdisciplinary project grant “Generation and transplantation of a bioengineered human bile duct” (F.S. and L.V.). L.V. lab is funded by the ERC proof of concept grant Relieve-Chol, by the ERC advanced grant New-Chol, the Cambridge University Hospitals National Institute for Health Research Biomedical Research Center and the core support grant from the Wellcome Trust and Medical Research Council to the Wellcome-Medical Research Council Cambridge Stem Cell Institute.
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Sampaziotis, F., Tysoe, O., Brevini, T., Vallier, L. (2019). Use of Biliary Organoids in Cholestasis Research. In: Vinken, M. (eds) Experimental Cholestasis Research. Methods in Molecular Biology, vol 1981. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9420-5_25
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DOI: https://doi.org/10.1007/978-1-4939-9420-5_25
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Publisher Name: Humana, New York, NY
Print ISBN: 978-1-4939-9419-9
Online ISBN: 978-1-4939-9420-5
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