Volume 31, No 3, Mar 2021

ISSN: 1001-0602 
EISSN: 1748-7838 2018 
impact factor 17.848* 
(Clarivate Analytics, 2019)

Volume 31 Issue 3, March 2021: 259-271   |  Open Access

ORIGINAL ARTICLES

Establishment of intestinal organoid cultures modeling injury-associated epithelial regeneration

Molong Qu¹ , Liang Xiong¹ , Yulin Lyu² , Xiannian Zhang³ , Jie Shen³ , Jingyang Guan¹ , Peiyuan Chai⁴ , Zhongqing Lin⁵ , Boyao Nie⁵ , Cheng Li^2,* , Jun Xu^1,* , Hongkui Deng^1,6,*

¹School of Basic Medical Sciences, State Key Laboratory of Natural and Biomimetic Drugs, Peking University Health Science Center and the MOE Key Laboratory of Cell Proliferation and Differentiation, College of Life Sciences, Peking-Tsinghua Center for Life Sciences, Peking University, Beijing 100191, China
²School of Life Sciences, Center for Bioinformatics, Center for Statistical Science, Peking University, Beijing 100871, China
³Department of Neurobiology, Capital Medical University, Beijing 100069, China
⁴Key Laboratory of Cell Proliferation and Differentiation of the Ministry of Education and State Key Laboratory of Membrane Biology, College of Life Sciences, Peking University, Beijing 100871, China
⁵Beijing Vitalstar Biotechnology Co., Ltd, Beijing 100000, China
⁶State Key Laboratory of Chemical Oncogenomics, School of Chemical Biology and Biotechnology, Peking University Shenzhen Graduate School, Shenzhen, Guangdong 518055, China
These authors contributed equally: Molong Qu, Liang Xiong, Yulin Lyu Correspondence: Cheng Li(cheng_li@pku.edu.cn)Jun Xu(jun_xu@bjmu.edu.cn)Hongkui Deng(hongkui_deng@pku.edu.cn)

The capacity of 3D organoids to mimic physiological tissue organization and functionality has provided an invaluable tool to model development and disease in vitro. However, conventional organoid cultures primarily represent the homeostasis of self-organizing stem cells and their derivatives. Here, we established a novel intestinal organoid culture system composed of 8 components, mainly including VPA, EPZ6438, LDN193189, and R-Spondin 1 conditioned medium, which mimics the gut epithelium regeneration that produces hyperplastic crypts following injury; therefore, these organoids were designated hyperplastic intestinal organoids (Hyper-organoids). Single-cell RNA sequencing identified different regenerative stem cell populations in our Hyper-organoids that shared molecular features with in vivo injury-responsive Lgr5+ stem cells or Clu+ revival stem cells. Further analysis revealed that VPA and EPZ6438 were indispensable for epigenome reprogramming and regeneration in Hyper-organoids, which functioned through epigenetically regulating YAP signaling. Furthermore, VPA and EPZ6438 synergistically promoted regenerative response in gut upon damage in vivo. In summary, our results demonstrated a new in vitro organoid model to study epithelial regeneration, highlighting the importance of epigenetic reprogramming that pioneers tissue repair.

https://doi.org/10.1038/s41422-020-00453-x

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