Stem/progenitor cells in murine mammary gland: isolation and functional characterization.
Stem/progenitor cells in murine mammary gland: isolation and functional characterization.
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DOI：
10.1007/978-1-61779-815-3_12
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发表时间：
2012-01-01
期刊：
影响因子：
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通讯作者：
Sun, Lu-Zhe
中科院分区：
文献类型：
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作者：
Bandyopadhyay, Abhik;Dong, Qiaoxiang;Sun, Lu-Zhe
成年功能性乳腺上皮干/祖细胞在乳腺中的存在以及最近通过特异性细胞表面标记鉴定出的可移植乳腺干细胞（MaSCs）富集部分彻底改变了描述乳腺干/祖细胞功能在乳腺发育和乳腺肿瘤发生中的作用的研究。在这一章中，我们已经详细介绍了从小鼠乳腺中分离乳腺上皮细胞的方法，通过荧光激活细胞分选（FACS）富集干/祖细胞组分，体外乳腺球培养，以及二维和三维培养中的分化测定。我们还描述了体内清除乳腺脂肪垫移植试验的详细方案，用于评估MaSC的再增殖活性，这表明它们在体内的多谱系分化和自我更新潜力，并被认为是功能性干细胞的“金标准”试验。
The presence of adult functional mammary epithelial stem/progenitor cells in mammary gland and recent identification of enriched fraction of transplantable mammary stem cells (MaSCs) through specific cell surface markers have revolutionized the study to delineate the role of mammary stem/progenitor functions in both mammary gland development and mammary tumorigenesis. In this chapter, we have described detailed methods of isolation of mammary epithelial cells from murine mammary glands, enrichment of stem/progenitor fractions by fluorescence-activated cell sorting (FACS), in vitro mammosphere culture, and differentiation assays in two- and three-dimensional culture. We have also described the detailed protocol of in vivo cleared mammary fat pad transplantation assay for the assessment of the MaSC repopulating activity, which indicates their multilineage differentiation and self-renewal potential in vivo and is considered the "gold standard" assay of functional stem cells.