Long-Term Expansion of Pancreatic Islet Organoids from Resident Procr+ Progenitors
Wang, Daisong; Wang, Jingqiang; Bai, Lanyue; Pan, Hong; Feng, Hua; Clevers, Hans; Zeng, Yi Arial
(2020) Cell, volume 180, issue 6, pp. 1198 - 1211.e19
(Article)
Abstract
It has generally proven challenging to produce functional β cells in vitro. Here, we describe a previously unidentified protein C receptor positive (Procr+) cell population in adult mouse pancreas through single-cell RNA sequencing (scRNA-seq). The cells reside in islets, do not express differentiation markers, and feature epithelial-to-mesenchymal transition characteristics. By
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genetic lineage tracing, Procr+ islet cells undergo clonal expansion and generate all four endocrine cell types during adult homeostasis. Sorted Procr+ cells, representing ∼1% of islet cells, can robustly form islet-like organoids when cultured at clonal density. Exponential expansion can be maintained over long periods by serial passaging, while differentiation can be induced at any time point in culture. β cells dominate in differentiated islet organoids, while α, δ, and PP cells occur at lower frequencies. The organoids are glucose-responsive and insulin-secreting. Upon transplantation in diabetic mice, these organoids reverse disease. These findings demonstrate that the adult mouse pancreatic islet contains a population of Procr+ endocrine progenitors.
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Keywords: Animals, Cell Culture Techniques/methods, Cell Differentiation/physiology, Cell Line, Cells, Cultured, Diabetes Mellitus, Experimental/metabolism, Endothelial Protein C Receptor/metabolism, Epithelial-Mesenchymal Transition/physiology, Female, Glucose/metabolism, Insulin Secretion, Insulin-Secreting Cells/cytology, Insulin/metabolism, Islets of Langerhans/cytology, Male, Mice, Mice, Inbred C57BL, Mice, Inbred ICR, Mice, Nude, Organoids/growth & development, Pancreas/cytology, Protein C/metabolism, Stem Cells/cytology, Procr, β cells, adult stem cells, organoid, pancreatic islets, General Biochemistry,Genetics and Molecular Biology, Research Support, Non-U.S. Gov't, Journal Article
ISSN: 0092-8674
Publisher: Cell Press
Note: Funding Information: Thanks to Ling Ge and Jun He in Cell Biology Core Facility of SIBCB for helps in electron microscopy experiments; to Dr. Changpeng Xin in Omics Core of Bio-Med Big Data Center, CAS-MPG Partner Institute for Computational Biology; to Drs. Zhiyong Liu and Chao Li of Institute of Neuroscience, Chinese Academy of Sciences; to Drs. Fuchou Tang, Li Li, and Lin Li of Peking University for help in single-cell sequencing experiments; to Drs. Chengran Xu and Xinxin Yu of Peking University for kindly providing the Ngn3-Cre mice and for help in single-cell sequencing experiments; to Biocytogen Co. for assistance in knockin mouse generation; to Dr. Chi Chung Hui of University of Toronto and Dr. Roel Nusse of Stanford University for helpful comments on the manuscript. This research was supported by grants from the Chinese Academy of Sciences ( XDB19020200 and XDA16020200 to Y.A.Z.); National Natural Science Foundation of China ( 31625020 , 31530045 , 31830056 , 31861163006 , and 31661143043 to Y.A.Z.); National Key Research and Development Program of China ( 2019YFA0802001 ); Shanghai Municipal Science and Technology Commission ( 17XD1404000 to Y.A.Z.); and CAS-NWO Programme Joint Research Project ( 153D31KYSB20150100 to Y.A.Z. and 40-41405-98-208 to H.C.). Funding Information: Thanks to Ling Ge and Jun He in Cell Biology Core Facility of SIBCB for helps in electron microscopy experiments; to Dr. Changpeng Xin in Omics Core of Bio-Med Big Data Center, CAS-MPG Partner Institute for Computational Biology; to Drs. Zhiyong Liu and Chao Li of Institute of Neuroscience, Chinese Academy of Sciences; to Drs. Fuchou Tang, Li Li, and Lin Li of Peking University for help in single-cell sequencing experiments; to Drs. Chengran Xu and Xinxin Yu of Peking University for kindly providing the Ngn3-Cre mice and for help in single-cell sequencing experiments; to Biocytogen Co. for assistance in knockin mouse generation; to Dr. Chi Chung Hui of University of Toronto and Dr. Roel Nusse of Stanford University for helpful comments on the manuscript. This research was supported by grants from the Chinese Academy of Sciences (XDB19020200 and XDA16020200 to Y.A.Z.); National Natural Science Foundation of China (31625020, 31530045, 31830056, 31861163006, and 31661143043 to Y.A.Z.); National Key Research and Development Program of China (2019YFA0802001); Shanghai Municipal Science and Technology Commission (17XD1404000 to Y.A.Z.); and CAS-NWO Programme Joint Research Project (153D31KYSB20150100 to Y.A.Z. and 40-41405-98-208 to H.C.). D.W. and Y.A.Z. designed the project and wrote the manuscript. D.W. performed most of the experiments including single-cell sequencing, lineage tracing, staining, in vitro culture, transplantation, and diabetic phenotype analysis. J.W. performed staining and diabetic phenotype analysis. D.W. and H.F. performed sequencing analysis. L.B. and H.P. performed genetic crosses. H.C. helped with project design and manuscript editing. Y.A.Z. and D.W. have a patent on islet organoid technology. H.C. is named as inventor on several organoid patents, but these are not related to the technology described here. All other authors declare no competing interests. Publisher Copyright: © 2020 Elsevier Inc. Copyright: Copyright 2021 Elsevier B.V., All rights reserved.
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