Prater Michael, Shehata Mona, Watson Christine J, Stingl John
Cancer Research, UK Cambridge Research Institute, Cambridge, UK.
Methods Mol Biol. 2013;946:395-409. doi: 10.1007/978-1-62703-128-8_25.
Evidence is emerging that the mouse mammary epithelium is arranged as a hierarchy that spans from stem cells to lineage-restricted progenitor cells to differentiated luminal and myoepithelial cells. The use of fluorescence-activated cell sorting (FACS) in combination with quantitative functional clonal assays represents a powerful tool for studying the properties of mouse mammary stem and progenitor cells. This chapter outlines the experimental procedures for generating single viable cell suspensions of mouse mammary epithelial cells, immunostaining cells for flow cytometry, in vitro assays for the detection and enumeration of mouse mammary progenitor cells, and in vivo assays for the detection and enumeration of mouse mammary stem cells.
越来越多的证据表明,小鼠乳腺上皮细胞呈层级排列,从干细胞到谱系受限的祖细胞,再到分化的管腔细胞和肌上皮细胞。将荧光激活细胞分选(FACS)与定量功能克隆分析相结合,是研究小鼠乳腺干细胞和祖细胞特性的有力工具。本章概述了生成小鼠乳腺上皮细胞单活细胞悬液的实验步骤、用于流式细胞术的细胞免疫染色、检测和计数小鼠乳腺祖细胞的体外分析,以及检测和计数小鼠乳腺干细胞的体内分析。