Identification, Isolation, and Culture of Intestinal Epithelial Stem Cells from Murine Intestine

Protocol
First Online:
Part of the Methods in Molecular Biology book series (MIMB, volume 879)

Abstract

The study of adult stem cell populations provides insight into the mechanisms that regulate tissue maintenance in normal physiology and many disease states. With an impressive rate of epithelial renewal driven by a pool of multipotent stem cells, the intestine is a particularly advantageous model system for the study of adult stem cells. Until recently, the isolation and in vitro study of intestinal epithelial stem cells (IESCs) was not possible due to the lack of biomarkers and culture techniques. However, advances in molecular characterization and culture of IESCs have made in vitro studies on this cell type amenable to most laboratories. The methods described in this chapter will allow the investigator to adapt newly established techniques toward downstream analysis of IESCs in vitro.

Key words

Intestinal epithelial stem cells Intestinal epithelial isolation Flow cytometry/fluorescence-activated cell sorting In vitro stemness assay Cell surface markers RT-PCR 
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Notes

Acknowledgments

We would like to thank Victoria Bali, Ph.D., for critical reading of the document. Additionally, we would like to acknowledge Jill Carrington, Ph.D., who to our knowledge was the first to coin the term “cryptoid.” The original work described in this chapter was funded by the National Institutes of Health, 1-K01-DK080181-01, the American Gastroenterological Association Research Scholar Award, the North Carolina Biotechnology Center Grant, and the UNC-Chapel Hill the Center for Gastrointestinal Biology and Disease, 5P30DK034987 (S.T. Magness).

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  1. 1.Department of Medicine, Division of Gastroenterology and Hepatology and Department of Cell and Molecular PhysiologyUniversity of North Carolina at Chapel HillChapel HillUSA
  2. 2.Departments of Medicine and Biomedical Engineering, Division of Gastroenterology and HepatologyUniversity of North Carolina and North Carolina State UniversityChapel HillUSA
  3. 3.Departments of Medicine and Biomedical Engineering, Division of Gastroenterology, and HepatologyUniversity of North Carolina and North Carolina State UniversityChapel HillUSA

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