Author:
Date:
2020-07-05
[Abstract] For years, the mammary gland serves as a perfect example to study the self-renew and differentiation of adult stem cells, and the regulatory mechanisms of these processes as well. To assess the function of given genes and/or other factors on stemness of mammary cells, several In vitro assays were developed, such as mammospheres formation assay, detection of stem cell markers by mRNA expression or flow cytometry and so on. However, the capacity of reconstruction of whole mount in the cleared fat pad of recipient female mice is a golden standard to estimate the stemness of the cells. Here we described a step-by-step protocol for in vivo mammary gland formation assay, including preparation of “cleared” recipients and mammary cells for implantation, the surgery process and ...
[摘要] [摘要 ] 多年来,乳腺一直是研究成体干细胞的自我更新和分化以及这些过程的调控机制的完美例证。为了评估给定的基因和/或其他因素对乳腺细胞的干性,有几个函数体外测定法开发出来,如微球体由mRNA表达形成试验,干细胞标志物的检测或流式细胞术等。然而,在雌性小鼠清除的脂肪垫中整个坐骨的重建能力是估计细胞干性的黄金标准。在这里,我们描述了体内的分步操作方案 乳腺形成测定,包括准备“清除”的受体和用于植入的乳腺细胞,手术过程以及如何评估实验结果。结合通过基因编辑和/或药物处理对乳腺细胞的操作,该方案在乳腺干细胞和乳腺发育的研究中可能非常有用。
[背景 ] 作为哺乳动物最典型的器官之一,乳腺(MG)是外分泌腺,负责泌乳。MG的发育受某些性激素的控制,这些激素的水平精确地调节了MG在不同发育阶段的结构,细胞组成和功能变化(Henigighausen and Robinson,2005)。许多遗传和环境因素都参与了乳腺干细胞的调控和MG的发育。为了研究这些因素的功能和机理,已经开发了几种方法,特别是用于评估乳腺细胞的干性。先前的研究表明,只有MG的基底细胞而非管腔细胞能够在受体雌性小鼠清除的脂肪垫中重建上皮树,这表明乳腺干细胞仅存在于基底谱系中(Van Keymeulen 等,2011)。 )。后来,包括我们在内的许多研究发现了乳腺干细胞的几种标志物(Prater ...
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