title: Systematically labeling developmental stage-specific genes for the study of pancreatic β-cell differentiation from human embryonic stem cells.
abstract: 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.
authors.affiliation: Shenzhen Stem Cell Engineering Laboratory, Key Laboratory of Chemical Genomics, Peking University Shenzhen Graduate School, Shenzhen, Guangdong 518055, China.
authors.forename: Haisong
authors.initials: H
authors.lastname: Liu
authors.affiliation: Shenzhen Stem Cell Engineering Laboratory, Key Laboratory of Chemical Genomics, Peking University Shenzhen Graduate School, Shenzhen, Guangdong 518055, China.
authors.forename: Huan
authors.initials: H
authors.lastname: Yang
authors.affiliation: Shenzhen Stem Cell Engineering Laboratory, Key Laboratory of Chemical Genomics, Peking University Shenzhen Graduate School, Shenzhen, Guangdong 518055, China.
authors.forename: Dicong
authors.initials: D
authors.lastname: Zhu
authors.affiliation: Shenzhen Stem Cell Engineering Laboratory, Key Laboratory of Chemical Genomics, Peking University Shenzhen Graduate School, Shenzhen, Guangdong 518055, China.
authors.forename: Xin
authors.initials: X
authors.lastname: Sui
authors.affiliation: Department of Biomedical Engineering, College of Engineering, Peking University, Beijing 100871, China.
authors.forename: Juan
authors.initials: J
authors.lastname: Li
authors.affiliation: Shenzhen Stem Cell Engineering Laboratory, Key Laboratory of Chemical Genomics, Peking University Shenzhen Graduate School, Shenzhen, Guangdong 518055, China.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Liang
authors.affiliation: Department of Hematopoietic Stem Cell Transplantation, 307 Hospital, Academy of Military Medicine Sciences, Beijing 100071, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Xu
authors.affiliation: The MOE Key Laboratory of Cell Proliferation and Differentiation, College of Life Sciences, Peking-Tsinghua Center for Life Sciences, Peking University, Beijing 100871, China.
authors.forename: Zeyu
authors.initials: Z
authors.lastname: Chen
authors.affiliation: The MOE Key Laboratory of Cell Proliferation and Differentiation, College of Life Sciences, Peking-Tsinghua Center for Life Sciences, Peking University, Beijing 100871, China.
authors.forename: Anzhi
authors.initials: A
authors.lastname: Yao
authors.affiliation: BGI-Shenzhen, Shenzhen, Guangdong 518083, China.
authors.forename: Long
authors.initials: L
authors.lastname: Zhang
authors.affiliation: BGI-Shenzhen, Shenzhen, Guangdong 518083, China.
authors.forename: Xi
authors.initials: X
authors.lastname: Zhang
authors.affiliation: The MOE Key Laboratory of Cell Proliferation and Differentiation, College of Life Sciences, Peking-Tsinghua Center for Life Sciences, Peking University, Beijing 100871, China.
authors.forename: Xing
authors.initials: X
authors.lastname: Yi
authors.affiliation: Shenzhen Stem Cell Engineering Laboratory, Key Laboratory of Chemical Genomics, Peking University Shenzhen Graduate School, Shenzhen, Guangdong 518055, China.
authors.forename: Meng
authors.initials: M
authors.lastname: Liu
authors.affiliation: Institute of Clinical Medical Sciences, China-Japan Friendship Hospital, Beijing 100029, China.
authors.forename: Shiqing
authors.initials: S
authors.lastname: Xu
authors.affiliation: Institute of Clinical Medical Sciences, China-Japan Friendship Hospital, Beijing 100029, China.
authors.forename: Wenjian
authors.initials: W
authors.lastname: Zhang
authors.affiliation: Department of Gynecology and Obstetrics, China-Japan Friendship Hospital, Beijing 100029, China.
authors.forename: Hua
authors.initials: H
authors.lastname: Lin
authors.affiliation: Department of Gynecology and Obstetrics, Beijing Renhe Hospital, Beijing 102600, China.
authors.forename: Lan
authors.initials: L
authors.lastname: Xie
authors.affiliation: Institute of Clinical Medical Sciences, China-Japan Friendship Hospital, Beijing 100029, China.
authors.forename: Jinning
authors.initials: J
authors.lastname: Lou
authors.affiliation: BGI-Shenzhen, Shenzhen, Guangdong 518083, China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Biomedical Engineering, College of Engineering, Peking University, Beijing 100871, China.
authors.forename: Jianzhong
authors.initials: J
authors.lastname: Xi
authors.affiliation: 1] Shenzhen Stem Cell Engineering Laboratory, Key Laboratory of Chemical Genomics, Peking University Shenzhen Graduate School, Shenzhen, Guangdong 518055, China [2] The MOE Key Laboratory of Cell Proliferation and Differentiation, College of Life Sciences, Peking-Tsinghua Center for Life Sciences, Peking University, Beijing 100871, China [3] Peking University Stem Cell Research Center, Department of Cell Biology, School of Basic Medical Sciences, Peking University Health Science Center, Beijing 100191, China.
authors.forename: Hongkui
authors.initials: H
authors.lastname: Deng
mesh_terms: D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics / Q000378:metabolism; D002454:Cell Differentiation; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D020869:Gene Expression Profiling; D017930:Genes, Reporter; D051542:Hepatocyte Nuclear Factor 3-beta / Q000235:genetics / Q000378:metabolism; D006801:Humans; D050417:Insulin-Secreting Cells / Q000166:cytology* / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D055760:SOXF Transcription Factors / Q000235:genetics / Q000378:metabolism; D017423:Sequence Analysis, RNA
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; D051542:Hepatocyte Nuclear Factor 3-beta; D008562:Membrane Glycoproteins; D009419:Nerve Tissue Proteins; D055760:SOXF Transcription Factors; C496502:FOXA2 protein, human; C498695:NEUROG3 protein, human; C109188:SOX17 protein, human; C582626:SUSD2 protein, human