Obtaining purified human intestinal epithelia for single-cell analysis and organoid culture.
Cell isolation
Clinical Protocol
Organoids
Single Cell
Stem Cells
Journal
STAR protocols
ISSN: 2666-1667
Titre abrégé: STAR Protoc
Pays: United States
ID NLM: 101769501
Informations de publication
Date de publication:
18 06 2021
18 06 2021
Historique:
entrez:
25
6
2021
pubmed:
26
6
2021
medline:
26
6
2021
Statut:
epublish
Résumé
Here, we describe protocols for the preparation and dissociation of human fetal and pediatric intestinal tissue to a high-viability epithelial single-cell suspension. This epithelium-enriched single-cell suspension can then be used to generate single-cell RNA sequencing data as well as to create human intestinal organoids from both the fetal and pediatric intestine. Finally, this protocol details the dissociation of the intestinal organoids for use in single-cell analysis or passaging of organoids. For complete details on the use and execution of this protocol, please refer to Elmentaite et al. (2020).
Identifiants
pubmed: 34169291
doi: 10.1016/j.xpro.2021.100597
pii: S2666-1667(21)00304-X
pmc: PMC8209738
doi:
Types de publication
Journal Article
Langues
eng
Pagination
100597Subventions
Organisme : Medical Research Council
ID : MR/T001917/1
Pays : United Kingdom
Informations de copyright
© 2021 The Authors.
Déclaration de conflit d'intérêts
The authors declare no competing interests.
Références
Gastroenterology. 2011 Nov;141(5):1762-72
pubmed: 21889923
Cell Stem Cell. 2013 Dec 5;13(6):734-44
pubmed: 24139758
Gut. 2019 Jan;68(1):49-61
pubmed: 29141958
Dev Cell. 2020 Dec 21;55(6):771-783.e5
pubmed: 33290721