ALDH as a marker for enriching tumorigenic human colonic stem cells

Anitha Shenoy, Elizabeth Butterworth, Emina H. Huang

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Aldehyde dehydrogenase (ALDH) can be used as a marker to isolate, propagate, and track normal and cancerous human colon stem cells. To determine their tumorigenic potential, tissues obtained from proximal (normal counterpart) and distal (cancerous) colon of colon cancer patients are implanted into NOD-SCID mice. In parallel, ALDHhigh and ALDHlow cells are isolated via Florescence Associated Cell Sorting (FACS) after the dissociation of distal and proximal colon tissues into a single-cell suspension. Flow cytometry for ALDHhigh and ALDHlow cells is possible with the ALDEFLUOR assay. Following cell sorting, ALDH-enriched cells are tested for their tumorigenic potential in vivo as xenografts. Owing to cancer stem cell properties, ALDHhigh cells could be propagated in vivo by serial passaging of the human tissue as xenografts and in vitro as suspension cultures called sphere cultures. In this unit, all the above-mentioned methods to isolate and propagate colon cancer stem cells using ALDH as a stem cell marker are described in detail.

Original languageEnglish (US)
Title of host publicationProgenitor Cells
Subtitle of host publicationMethods and Protocols
PublisherHumana Press Inc.
Pages373-385
Number of pages13
ISBN (Print)9781617799792
DOIs
StatePublished - 2012
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume916
ISSN (Print)1064-3745

Keywords

  • Aldehyde dehydrogenase
  • Colon cancer
  • Fluorescent activated cell sorting
  • Sphere cultures
  • Xenograft

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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