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Author:
Date:
2014-04-05
[Abstract] The mammary epithelium consists of multiple phenotypically and functionally distinct cell populations, which are organized as a hierarchy of stem cells, progenitors and terminally differentiated cells. Identification of the mechanisms regulating the growth and differentiation of mammary stem and progenitor cells is of great interest not only to better understand the mammary gland development but also to clarify the origins of breast cancer, as these cells seem to be the likely targets of malignant transformation within the mammary epithelium. Hence, a variety of approaches have been developed ...
[摘要] 乳腺上皮由多个表型和功能不同的细胞群组成，其被组织为干细胞，祖细胞和终末分化细胞的层次结构。鉴定调节乳腺干细胞和祖细胞的生长和分化的机制是非常感兴趣的，不仅是为了更好地理解乳腺发育，而且澄清乳腺癌的起源，因为这些细胞似乎是恶性转化的可能靶标在乳腺上皮内。因此，已经开发了多种方法用于定量和研究这些特定的乳腺细胞亚群。
鉴于它们的高增殖能力，乳腺祖细胞能够在低密度培养物中体外形成集落。在这里我们描述如何执行三维（3D）乳房集落形成细胞（Ma-CFC）测定，一种体外功能测定，适用于检测和分析无饲养层的乳腺祖细胞培养条件。
简单来说，这个协议涉及以克隆密度将乳腺单细胞接种到半固体基质（Matrigel）上，从而允许乳腺祖细胞增殖并产生离散的3D集落。所得菌落的数量和细胞组成将根据祖细胞的频率和分化潜能而分别变化。
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