| A 3D Culture System of Human Immortalized Myometrial Cells
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Author:
Date:
2016-10-20
[Abstract] Myometrium forms the middle layer of the uterus and is mainly composed of the smooth muscle cells. The cells in vitro are usually grown in a single layer (2-dimensional; 2D) format, whereas in vivo cells are structured in an extracellular matrix scaffolding that allows the cells to communicate and respond to environmental cues. We have developed human myometrium and leiomyoma 3-dimensional (3D) culture, wherein the cells retain their molecular characteristics and respond to environmental cues (Malik and Catherino, 2012; Malik et al., 2014).
[摘要] 子宫肌层形成子宫的中间层，主要由平滑肌细胞组成。细胞在体外通常生长在单层（2维; 2D）格式中，而体内细胞在细胞外基质支架中结构化，其允许细胞沟通和响应环境线索。我们已经开发了人子宫肌瘤和平滑肌瘤3维（3D）培养物，其中细胞保留其分子特征并响应环境线索（Malik和Catherino，2012; Malik等人，2014）。 br /> [背景] 在过去十年中，随着更多实验室从使用人工2D格式的细胞培养转移到3D细胞培养模型系统，观察到一定的转变，其中细胞生长在允许它们附着并获得更生理结构的基质中。该模型系统为细胞提供更自然的分化状态，并且培养的细胞在体内形成组织样环境。这是一个详细的协议为myometrium 3D细胞培养生长胶原-1矩阵，修改从Malik和Catherino（2012年）。
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| 3D Mammary Colony-Forming Cell Assay
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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 for quantifying and studying these specific mammary cell subsets.
Given their high proliferative capacity, mammary progenitor cells are able to form colonies in vitro ...
[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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