Introducing the Stem Cell ASCL2 Reporter STAR into Intestinal Organoids

Maria C Heinz; Koen C Oost; Hugo J G Snippert

doi:10.1016/j.xpro.2020.100126

Introducing the Stem Cell ASCL2 Reporter STAR into Intestinal Organoids

STAR Protoc. 2020 Oct 8;1(3):100126. doi: 10.1016/j.xpro.2020.100126. eCollection 2020 Dec 18.

Authors

Maria C Heinz^{1

2}, Koen C Oost^{1

2}, Hugo J G Snippert^{1

2}

Affiliations

¹ Molecular Cancer Research, Center for Molecular Medicine, University Medical Center Utrecht, University Utrecht, Utrecht, The Netherlands.
² Oncode Institute, Utrecht, The Netherlands.

Abstract

Patient-derived organoids maintain functional and phenotypic characteristics of the original tissue such as cell-type diversity. Here, we provide protocols on how to label intestinal (cancer) stem cells by integrating the stem cell ASCL2 reporter (STAR) into human and mouse genomes via two different strategies: (1) lentiviral transduction or (2) transposon-based integration. Organoid technology, in combination with the user-friendly nature of STAR, will facilitate basic research in human and mouse adult stem cell biology. For complete details on the use and execution of this protocol, please refer to Oost et al. (2018).

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adult Stem Cells / metabolism
Animals
Basic Helix-Loop-Helix Transcription Factors / genetics
Humans
Intestinal Mucosa / diagnostic imaging
Intestines / cytology
Mice
Organoids / growth & development
Organoids / metabolism*
Staining and Labeling / methods*
Stem Cells / metabolism

Substances

ASCL2 protein, human
Basic Helix-Loop-Helix Transcription Factors