Establishment and Culture of Human Intestinal Organoids Derived from Adult Stem Cells
Pleguezuelos-Manzano, Cayetano; Puschhof, Jens; van den Brink, Stieneke; Geurts, Veerle; Beumer, Joep; Clevers, Hans
(2020) Current protocols in immunology, volume 130, issue 1
(Article)
Abstract
Human intestinal organoids derived from adult stem cells are miniature ex vivo versions of the human intestinal epithelium. Intestinal organoids are useful tools for the study of intestinal physiology as well as many disease conditions. These organoids present numerous advantages compared to immortalized cell lines, but working with them requires
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dedicated techniques. The protocols described in this article provide a basic guide to establishment and maintenance of human intestinal organoids derived from small intestine and colon biopsies. Additionally, this article provides an overview of several downstream applications of human intestinal organoids.
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Keywords: adult stem cells, human intestinal organoids, organoid cryopreservation, organoid culture establishment, organoid differentiation, organoid immunofluorescence, organoid passage, single-cell clonal organoid culture, specialized organoid reagents, Immunology
ISSN: 1934-3671
Publisher: John Wiley & Sons Inc.
Note: Funding Information: We thank M. Bannier‐Hélaouët for critical reading of the manuscript and comments. Development of this method was supported by CRUK grant OPTIMISTICC (C10674/A27140) (C.P.‐M., J.P.), the Gravitation projects CancerGenomiCs.nl, and the Netherlands Organ‐on‐Chip Initiative (024.003.001) from the Netherlands Organisation for Scientific Research (NWO) funded by the Ministry of Education, Culture and Science of the government of the Netherlands (C.P.‐M., J.P.), the Oncode Institute (partly financed by the Dutch Cancer Society), the European Research Council under ERC Advanced Grant Agreement no. 67013 (J.P., H.C.) and NETRF/Petersen Accelerator (J.B.). Funding Information: We thank M. Bannier-H?laou?t for critical reading of the manuscript and comments. Development of this method was supported by CRUK grant OPTIMISTICC (C10674/A27140) (C.P.-M., J.P.), the Gravitation projects CancerGenomiCs.nl, and the Netherlands Organ-on-Chip Initiative (024.003.001) from the Netherlands Organisation for Scientific Research (NWO) funded by the Ministry of Education, Culture and Science of the government of the Netherlands (C.P.-M., J.P.), the Oncode Institute (partly financed by the Dutch Cancer Society), the European Research Council under ERC Advanced Grant Agreement no. 67013 (J.P., H.C.) and NETRF/Petersen Accelerator (J.B.). Publisher Copyright: © 2020 The Authors.
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