Volume 24, No 10, Oct 2014

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

Volume 24 Issue 10, October 2014: 1181-1200

ORIGINAL ARTICLES

Systematically labeling developmental stage-specific genes for the study of pancreatic β-cell differentiation from human embryonic stem cells

Haisong Liu^1,*, Huan Yang^1,*, Dicong Zhu^1,*, Xin Sui¹, Juan Li², Zhen Liang¹, Lei Xu³, Zeyu Chen⁴, Anzhi Yao⁴, Long Zhang⁵, Xi Zhang⁵, Xing Yi⁴, Meng Liu¹, Shiqing Xu⁶, Wenjian Zhang⁶, Hua Lin⁷, Lan Xie⁸, Jinning Lou⁶, Yong Zhang⁵, Jianzhong Xi² and Hongkui Deng^1,4,9

¹Shenzhen Stem Cell Engineering Laboratory, Key Laboratory of Chemical Genomics, Peking University Shenzhen Graduate School, Shenzhen, Guangdong 518055, China
²Department of Biomedical Engineering, College of Engineering, Peking University, Beijing 100871, China
³Department of Hematopoietic Stem Cell Transplantation, 307 Hospital, Academy of Military Medicine Sciences, Beijing 100071, China
⁴The MOE Key Laboratory of Cell Proliferation and Differentiation, College of Life Sciences, Peking-Tsinghua Center for Life Sciences, Peking University, Beijing 100871, China
⁵BGI-Shenzhen, Shenzhen, Guangdong 518083, China
⁶Institute of Clinical Medical Sciences, China-Japan Friendship Hospital, Beijing 100029, China
⁷Department of Gynecology and Obstetrics, China-Japan Friendship Hospital, Beijing 100029, China
⁸Department of Gynecology and Obstetrics, Beijing Renhe Hospital, Beijing 102600, China
⁹Peking University Stem Cell Research Center, Department of Cell Biology, School of Basic Medical Sciences, Peking University Health Science Center, Beijing 100191, China Correspondence: Hongkui Deng, E-mail: hongkui_deng@pku.edu.cn; Jianzhong Xi,(xi@coe.pku.edu.cn)

The applications of human pluripotent stem cell (hPSC)-derived cells in regenerative medicine has encountered a long-standing challenge: how can we efficiently obtain mature cell types from hPSCs? Attempts to address this problem are hindered by the complexity of controlling cell fate commitment and the lack of sufficient developmental knowledge for guiding hPSC differentiation. Here, we developed a systematic strategy to study hPSC differentiation by labeling sequential developmental genes to encompass the major developmental stages, using the directed differentiation of pancreatic β cells from hPSCs as a model. We therefore generated a large panel of pancreas-specific mono- and dual-reporter cell lines. With this unique platform, we visualized the kinetics of the entire differentiation process in real time for the first time by monitoring the expression dynamics of the reporter genes, identified desired cell populations at each differentiation stage and demonstrated the ability to isolate these cell populations for further characterization. We further revealed the expression profiles of isolated NGN3-eGFP+ cells by RNA sequencing and identified sushi domain-containing 2 (SUSD2) as a novel surface protein that enriches for pancreatic endocrine progenitors and early endocrine cells both in human embryonic stem cells (hESC)-derived pancreatic cells and in the developing human pancreas. Moreover, we captured a series of cell fate transition events in real time, identified multiple cell subpopulations and unveiled their distinct gene expression profiles, among heterogeneous progenitors for the first time using our dual reporter hESC lines. The exploration of this platform and our new findings will pave the way to obtain mature β cells in vitro.

10.1038/cr.2014.118

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