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Book chapter
Open access
English

Studying Stem Cell Biology in Intact and Whole-Body Regenerating Hydra by Flow Cytometry

PublisherNew York : Humana Press
Collection
  • Methods in molecular biology; 2450
Publication date2022-03-31
First online date2022-04-01
Abstract

The freshwater Hydra polyp is a versatile model to study whole-body regeneration from a developmental as well as a cellular point of view. The outstanding regenerative capacities of Hydra are based on its three populations of adult stem cells located in the central body column of the animal. There, these three populations, gastrodermal epithelial, epidermal epithelial, and interstitial, continuously cycle in homeostatic conditions, and their activity is locally regulated after mid-gastric bisection. Moreover, they present an unusual cycling behavior with a short G1 phase and a pausing in G2. This particular cell cycle has been studied for a long time with classical microscopic methods. We describe here two flow cytometry methods that provide accurate and reproducible quantitative data to monitor cell cycle regulation in homeostatic and regenerative contexts. We also present a cell sorting procedure based on flow cytometry, whereby stem cells expressing a fluorescent reporter protein in transgenic lines can be enriched for use in applications such as transcriptomic, proteomic, or cell cycle analysis.

eng
Keywords
  • Hydra
  • Cell cycle
  • Adult stem cells
  • Epithelial stem cells
  • Interstitial stem cells
  • Flow cytometry
  • GFP cell sorting
  • Click-iT EdU labeling of regenerating animals
  • Cell cycle analysis
Research group
Citation (ISO format)
BUZGARIU, Wanda Christa, AUBRY-LACHAINAYE, Jean-Pierre, GALLIOT, Brigitte. Studying Stem Cell Biology in Intact and Whole-Body Regenerating Hydra by Flow Cytometry. In: Whole-Body Regeneration: Methods and Protocols. New York : Humana Press, 2022. p. 373–398. (Methods in molecular biology) doi: 10.1007/978-1-0716-2172-1_20
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Creation04/02/2022 12:22:00 PM
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