Henseleit Korinna D, Wheeler Ann P, Warnes Gary, Braun Kristin M
Centre for Cutaneous Research, Blizard Institute, Barts and the London School of Medicine and Dentistry, Queen Mary, University of London, London, UK.
Methods Mol Biol. 2012;916:243-61. doi: 10.1007/978-1-61779-980-8_19.
The epidermis and its appendages, the hair follicle and sebaceous gland, have the capacity to constantly regenerate throughout adult life. Postnatal hair follicles undergo a cyclic mode of tissue homeostasis, defined by periods of growth, degeneration, and rest. A multipotent population of stem cells residing within the hair follicle bulge not only generates the hair lineages during each hair cycle, but also transiently contributes to the repair of epidermis following wounding. In this chapter, we provide methods for identifying epidermal stem cells and investigating their proliferative and apoptotic characteristics. We introduce whole-mount and flow cytometry techniques, which complement each other by permitting visualization of the epidermal stem cell compartment in situ and assessment of the phenotype of purified cells. These techniques can easily be adapted to characterize novel putative epidermal stem or progenitor cell populations. By applying whole-mount and flow cytometry techniques to characterize normal and genetically modified mice with skin defects, we expect to learn more about the factors that regulate stem cell self-renewal and differentiation.
表皮及其附属器，即毛囊和皮脂腺，在成年期具有持续再生的能力。出生后的毛囊经历一种组织稳态的循环模式，由生长、退化和休止期定义。位于毛囊隆突部的多能干细胞群体不仅在每个毛发周期产生毛发谱系，而且在受伤后还短暂地参与表皮的修复。在本章中，我们提供了鉴定表皮干细胞并研究其增殖和凋亡特性的方法。我们介绍了整装和流式细胞术技术，这两种技术相互补充，通过原位可视化表皮干细胞区室和评估纯化细胞的表型。这些技术可以很容易地用于表征新的假定表皮干细胞或祖细胞群体。通过应用整装和流式细胞术技术来表征有皮肤缺陷的正常和转基因小鼠，我们期望更多地了解调节干细胞自我更新和分化的因素。
