Morphogenesis and oncogenesis of MCF-10A mammary epithelial acini grown in three-dimensional basement membrane cultures
Morphogenesis and oncogenesis of MCF-10A mammary epithelial acini grown in three-dimensional basement membrane cultures
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DOI：
10.1016/s1046-2023(03)00032-x
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发表时间：
2003-07-01
期刊：
影响因子：
4.8
通讯作者：
Brugge, JS
中科院分区：
文献类型：
--
作者：
Debnath, J;Muthuswamy, SK;Brugge, JS
MCF-10A乳腺上皮细胞在重构基底膜上的三维培养导致形成极化的、生长停滞的腺泡样球状体，其概括了体内腺体结构的几个方面。导入MCF-10A细胞的癌基因破坏了这种形态发生过程，并引起不同的形态表型。最近的研究分析了在不同癌基因中观察到的表型异质性的机制基础（例如，ErbB 2、细胞周期蛋白D1）已经说明了这种三维培养系统在模拟癌基因的生物活性中的效用，特别是关于它们在癌形成的早期方面破坏上皮结构的能力。在这里，我们提供了一系列培养MCF-10A细胞的方案，通过逆转录病毒感染建立表达感兴趣基因的稳定库，以及在三维基底膜培养中生长和分析MCF-10A细胞。(C)2003 Elsevier Science（美国）。All rights reserved.
The three-dimensional culture of MCF-10A mammary epithelial cells on a reconstituted basement membrane results in formation of polarized, growth-arrested acini-like spheroids that recapitulate several aspects of glandular architecture in vivo. Oncogenes introduced into MCF-10A cells disrupt this morphogenetic process, and elicit distinct morphological phenotypes. Recent studies analyzing the mechanistic basis for phenotypic heterogeneity observed among different oncogenes (e.g., ErbB2, cyclin D1) have illustrated the utility of this three-dimensional culture system in modeling the biological activities of cancer genes, particularly with regard to their ability to disrupt epithelial architecture during the early aspects of carcinoma formation. Here we provide a collection of protocols to culture MCF-10A cells, to establish stable pools expressing a gene of interest via retroviral infection, as well as to grow and analyze MCF-10A cells in three-dimensional basement membrane culture. (C) 2003 Elsevier Science (USA). All rights reserved.