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Methods Mol Biol. 2016 Oct 5. [Epub ahead of print]

Isolation and Culture of Adult Intestinal, Gastric, and Liver Organoids for Cre-recombinase-Mediated Gene Deletion.

Author information

1
Molecular Oncology Laboratory, University of Melbourne, Melbourne, VIC, 3000, Australia.
2
Victorian Infectious Diseases Reference Laboratory, Doherty Institute, 792 Elizabeth Street, Melbourne, VIC, 3000, Australia.
3
Molecular Oncology Laboratory, University of Melbourne, Melbourne, VIC, 3000, Australia. evincan@unimelb.edu.au.
4
Victorian Infectious Diseases Reference Laboratory, Doherty Institute, 792 Elizabeth Street, Melbourne, VIC, 3000, Australia. evincan@unimelb.edu.au.
5
School of Biomedical Sciences, Curtin University, Perth, WA, 6845, Australia. evincan@unimelb.edu.au.

Abstract

The discovery of Lgr5 as a marker of adult stem cells meant that stem cell populations could be purified and studied in isolation. Importantly, when cultured under the appropriate conditions these stem cells form organoids in tissue culture that retain many features of the tissue of origin. The organoid cultures are accessible to genetic and biochemical manipulation, bridging the gap between in vivo mouse models and conventional tissue culture. Here we describe robust protocols to establish organoids from gastrointestinal tissues (stomach, intestine, liver) and Cre-recombinase mediated gene manipulation in vitro.

KEYWORDS:

3D organoid culture; Cre-recombinase; Gastric organoids; Intestinal organoids; Lgr5; Liver organoids

PMID:
27704362
DOI:
10.1007/7651_2016_14

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