Establishment of Human- and Mouse-Derived Gastric Primary Epithelial Cell Monolayers from Organoids

Emma Teal; Nina Bertaux-Skeirik; Jayati Chakrabarti; Loryn Holokai; Yana Zavros

doi:10.1007/978-1-4939-8600-2_15

Establishment of Human- and Mouse-Derived Gastric Primary Epithelial Cell Monolayers from Organoids

Methods Mol Biol. 2018:1817:145-155. doi: 10.1007/978-1-4939-8600-2_15.

Authors

Emma Teal¹, Nina Bertaux-Skeirik¹, Jayati Chakrabarti¹, Loryn Holokai¹, Yana Zavros²

Affiliations

¹ Department of Pharmacology and Systems Physiology, University of Cincinnati, Cincinnati, OH, USA.
² Department of Pharmacology and Systems Physiology, University of Cincinnati, Cincinnati, OH, USA. yana.zavros@uc.edu.

PMID: 29959711
DOI: 10.1007/978-1-4939-8600-2_15

Abstract

Organoid cultures generated from gastrointestinal tissues have been an invaluable advancement for in vitro studies of physiological function and disease. Here we present a comprehensive protocol for the establishment and culture of human- and mouse-derived 3-dimensional gastric organoids transferred to 2-dimensional gastric epithelial cell monolayers. We introduce two methods that include the establishment of monolayers from: (1) intact organoids, and (2) single cells dissociated from intact organoids.

Keywords: Gastric organoids; Monolayer; Regeneration; Repair; Scratch wound assay; Stem cells.

Publication types

Research Support, N.I.H., Extramural

MeSH terms

Animals
Epithelial Cells / cytology
Gastric Mucosa / cytology*
Humans
Mice
Organoids / cytology*
Primary Cell Culture / methods*

Grants and funding

U19 AI116491/AI/NIAID NIH HHS/United States