Directed differen a on of human pluripotent stem cells into hepatic tissue with gallbladder and bile ducts organoids in vitro

Publisher:
Oxford University Press (OUP)
Publication Type:
Conference Proceeding
Citation:
Annals of Oncology, 2019, 30 (4)
Issue Date:
2019-07-02
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Background: Liver transplantation is an effective treatment for patients with end-stage liver diseases such as liver cancer. However, liver transplant is heavily restricted by the shortage of donor organs. To date, organoids have been generated for a number of organs from human stem cells. Here we establish a robust and efficient process to direct the differentiation of human pluripotent stem cells (PSCs) into hepatic tissue with gallbladder and bile ducts in vitro using MIX medium. Methods: At differentiation stages I and II (day 1–15), 5%-25% of mTeSR TM culture medium was added to hepatic differentiation medium to induce endodermal and mesodermal commitment and thereafter hepatic and biliary co-differentiation. At stage III (day 15–45), 10% cholesterol + MIX was added to the maturation medium to promote the formation and maturation of the hepatobiliary organoids. Phenotypes and functions of organoids were determined by specific markers and multiple functional assays both in vitro and in vivo. The differentiation of PSCs was in 2-dimensional (2D) monolayer cultures rather than 3D cultures. Results: The resulting hepatic tissue consisted of a polygonal hepatocyte that was patterned into liver parenchyma structures and gallbladder and bile ducts organoids contained cholangiocytes. The hepatocytes of liver parenchyma-like tissue exhibit characteristics of mature hepatocytes. Particularly, these cells could produced bile acid, which were stored in the gallbladder and bile duct organoids. After transplantation into an immunedeficient mouse model, liver parenchyma–like tissue with gallbladder and bile duct organoids could be survival. Conclusion(s): Our findings can provide a powerful system not only for studying the molecular basis of human liver and gall disease in vitro but also for generating functional liver for eventual transplantation-based therapy for liver diseases.
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