Establishment and Culture of Human Intestinal Organoids Derived from Adult Stem Cells.
Adult Stem Cells
/ cytology
Biomarkers
Biopsy
Cell Culture Techniques
Cell Differentiation
Cell Separation
/ methods
Cells, Cultured
Colon
/ cytology
Cryopreservation
/ methods
Culture Media, Conditioned
Fluorescent Antibody Technique
Humans
Immunohistochemistry
Immunophenotyping
Intestinal Mucosa
/ cytology
Intestine, Small
/ cytology
Organoids
/ cytology
Tissue Culture Techniques
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
Journal
Current protocols in immunology
ISSN: 1934-368X
Titre abrégé: Curr Protoc Immunol
Pays: United States
ID NLM: 9101651
Informations de publication
Date de publication:
09 2020
09 2020
Historique:
entrez:
17
9
2020
pubmed:
18
9
2020
medline:
16
6
2021
Statut:
ppublish
Résumé
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 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. © 2020 The Authors. Basic Protocol 1: Establishment of human small intestine and colon organoid cultures from fresh biopsies Basic Protocol 2: Mechanical splitting, passage, and expansion of human intestinal organoids Alternate Protocol: Differentiation of human intestinal organoids Basic Protocol 3: Cryopreservation and thawing of human intestinal organoids Basic Protocol 4: Immunofluorescence staining of human intestinal organoids Basic Protocol 5: Generation of single-cell clonal intestinal organoid cultures Support Protocol 1: Production of Wnt3A conditioned medium Support Protocol 2: Production of Rspo1 conditioned medium Support Protocol 3: Extraction of RNA from intestinal organoid cultures.
Identifiants
pubmed: 32940424
doi: 10.1002/cpim.106
pmc: PMC9285512
doi:
Substances chimiques
Biomarkers
0
Culture Media, Conditioned
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
e106Subventions
Organisme : Cancer Research UK
ID : C10674/A27140
Pays : United Kingdom
Informations de copyright
© 2020 The Authors.
Références
Nature. 2016 Feb 18;530(7590):340-3
pubmed: 26863187
Nature. 2020 Apr;580(7802):269-273
pubmed: 32106218
Nat Cell Biol. 2018 Aug;20(8):909-916
pubmed: 30038251
Gastroenterology. 2019 Feb;156(3):562-576
pubmed: 30476497
Cell. 2013 Jul 18;154(2):274-84
pubmed: 23870119
Cell. 2015 May 7;161(4):933-45
pubmed: 25957691
Cell. 2018 Sep 6;174(6):1586-1598.e12
pubmed: 30100188
Nat Commun. 2019 Oct 8;10(1):4571
pubmed: 31594944
Cell Stem Cell. 2020 Nov 5;27(5):840-851.e6
pubmed: 32818433
Curr Protoc Immunol. 2020 Sep;130(1):e106
pubmed: 32940424
Nat Rev Immunol. 2020 May;20(5):279-293
pubmed: 31853049
Gastroenterology. 2018 Jul;155(1):130-143.e15
pubmed: 29604290
Cell Stem Cell. 2013 Dec 5;13(6):653-8
pubmed: 24315439
Sci Rep. 2019 May 7;9(1):7029
pubmed: 31065051
Nature. 2016 Oct 13;538(7624):260-264
pubmed: 27698416
Nature. 2009 May 14;459(7244):262-5
pubmed: 19329995
Nature. 2020 Jan;577(7789):254-259
pubmed: 31853059
Nat Protoc. 2015 Oct;10(10):1474-85
pubmed: 26334867
Gastroenterology. 2011 Nov;141(5):1762-72
pubmed: 21889923
Cell. 2016 Jun 16;165(7):1586-1597
pubmed: 27315476
Cell. 2020 Jun 11;181(6):1291-1306.e19
pubmed: 32407674
Nat Med. 2013 Jul;19(7):939-45
pubmed: 23727931
Cell. 2004 Mar 19;116(6):883-95
pubmed: 15035989
Nature. 2015 May 7;521(7550):43-7
pubmed: 25924068
Nat Rev Cancer. 2018 Jul;18(7):407-418
pubmed: 29692415
Cell Stem Cell. 2018 Dec 6;23(6):787-793.e6
pubmed: 30526881
Nature. 2017 May 11;545(7653):234-237
pubmed: 28467818
Sci Rep. 2020 Feb 26;10(1):3535
pubmed: 32103108
Cell Stem Cell. 2020 Apr 2;26(4):503-510.e7
pubmed: 32084388
J Clin Invest. 2014 Jan;124(1):328-37
pubmed: 24292712