Department of Genetics and Evolution, iGE3, Faculty of Sciences, University of Geneva, Geneva, Switzerland.
Flow Cytometry Facility, Faculty of Medicine, University of Geneva, Geneva, Switzerland.
Methods Mol Biol. 2022;2450:373-398. doi: 10.1007/978-1-0716-2172-1_20.
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.
淡水水螅水螅体是一种多功能的模型，可从发育和细胞的角度研究全身再生。水螅具有出色的再生能力，这基于其位于动物中央体柱中的三个成年干细胞群体。在那里，这三个群体——胃上皮、表皮上皮和间质——在稳态条件下不断循环，并且在胃中部二分之后其活性受到局部调节。此外，它们表现出一种不寻常的循环行为，具有短的 G1 期和 G2 期暂停。这种特殊的细胞周期已经使用经典的显微镜方法进行了长时间的研究。我们在这里描述了两种流式细胞术方法，这些方法提供了准确和可重复的定量数据，可用于监测稳态和再生环境中的细胞周期调控。我们还介绍了一种基于流式细胞术的细胞分选程序，通过该程序，可以富集表达荧光报告蛋白的干细胞，用于转录组学、蛋白质组学或细胞周期分析等应用。
