Abstract
We report the generation of human ESC-derived, expandable hepatic organoids (hEHOs) using our newly established method with wholly defined (serum-free, feeder free) media. The hEHOs stably maintain phenotypic features of bipotential liver stem/progenitor cells that can differentiate into functional hepatocytes or cholangiocytes. The hEHOs can expand for 20 passages enabling large scale expansion to cell numbers requisite for industry or clinical programs. The cells from hEHOs display remarkable repopulation capacity in injured livers of FRG mice following transplantation, and they differentiate in vivo into mature hepatocytes. If implanted into the epididymal fat pads of immune-deficient mice, they do not generate non-hepatic lineages and have no tendency to form teratomas. We further develop a derivative model by incorporating human fetal liver mesenchymal cells (hFLMCs) into the hEHOs, referred to as hFLMC/hEHO, which can model alcoholic liver disease-associated pathophysiologic changes, including oxidative stress generation, steatosis, inflammatory mediators release and fibrosis, under ethanol treatment. Our work demonstrates that the hEHOs have considerable potential to be a novel, ex vivo pathophysiological model for studying alcoholic liver disease as well as a promising cellular source for treating human liver diseases.
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Acknowledgements
We thank Drs Zhigui Zeng, and Zhijun Zhu for their help with clinical samples; Drs Lola Reid and Xin Wang for critical review. Dr Xin Chang for TEM experiments; Mr Zhimin Li and Chuanwen Wang for bioinformatics analysis. This work was supported by the National Natural Science Foundations of China (No. 81730052), the Interdisciplinary Cooperation Project of Beijing Nova Program (Z1811100006218127), the National Major Scientific and Technological Special Project for “Significant New Drugs Development” (2018ZX09711003–001–002), the National Key Research and Development Program of China (No. 2016YFC1101305), the Science and Technology Planning Project of Guangdong China (2015A050502023), the Guangdong Province Science and Technology Program (2018KJYZ021) and Science and Technology Program of Guangzhou, China (STPG; 2016201604030054).
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Y.W. and S.W. conceived and designed the project. S.W. and X.W. conducted most of the experiments. Y.W., S.W., and X.W. wrote and edited the manuscript. Z.T., Y.S., and M.C. contributed to studies with cell culture, IF and tissue histology. J.L. helped with HCA experiments. F.Y. helped with in vivo transplantation experiments. J.C., T.C., C.L., and J.H. reviewed the final version of the manuscript.
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These authors are co-senior authors: Jie Hu, Yunfang Wang
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Wang, S., Wang, X., Tan, Z. et al. Human ESC-derived expandable hepatic organoids enable therapeutic liver repopulation and pathophysiological modeling of alcoholic liver injury. Cell Res 29, 1009–1026 (2019). https://doi.org/10.1038/s41422-019-0242-8
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DOI: https://doi.org/10.1038/s41422-019-0242-8
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