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Obtaining purified human intestinal epithelia for single-cell analysis and organoid culture.

ABSTRACT: 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).

SUBMITTER: Ross ADB

PROVIDER: S-EPMC8209738 | biostudies-literature | 2021 Jun

REPOSITORIES: biostudies-literature

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Obtaining purified human intestinal epithelia for single-cell analysis and organoid culture.

Ross Alexander D B ADB Perrone Francesca F Elmentaite Rasa R Teichmann Sarah A SA Zilbauer Matthias M

STAR protocols 20210612 2

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 organoi ...[more]

PMID: 34169291

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