Three-dimensional Mammary Epithelial Cell Morphogenesis Model for Analysis of TGFß Signaling.

Three-dimensional Mammary Epithelial Cell Morphogenesis Model for Analysis of TGFß Signaling.
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用于分析 TGFα 信号传导的三维乳腺上皮细胞形态发生模型。

DOI:
10.1007/978-1-4939-2966-5_7
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发表时间:
2016
期刊:
Methods in molecular biology (Clifton, N.J.)
影响因子:
--
通讯作者:
Luo,Kunxin
Luo,Kunxin
中科院分区:
--
文献类型:
--
作者:
Rashidian,Juliet;Luo,Kunxin

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在富含层粘连蛋白的细胞外基质中培养乳腺上皮细胞(三维或3D培养)提供了优于常规二维(2D)组织培养系统的显著优势,因为它考虑了细胞外基质(ECM)微环境对乳腺上皮细胞增殖、存活和分化的影响。当在3D培养物中生长时,未转化的乳腺上皮细胞经历形态发生以形成多细胞和极化的腺泡样结构,其在功能上模拟妊娠乳腺中的分化的肺泡。这一过程受到许多生长因子和细胞因子的调节。转化生长因子-β(TGF β)是一种多能细胞因子,调节发育和肿瘤发生的多个方面。除了其对上皮细胞增殖、存活和分化的作用外,它也是细胞-基质相互作用的有效调节剂。因此,3D培养模型可以概括复杂的体内上皮细胞微环境,并允许我们充分评估TGF β信号传导在正常和癌细胞行为的多个方面中的作用。在本章中,我们提供了详细的协议,在3D基质胶生长乳腺上皮细胞的信号通路的分析。
Culturing mammary epithelial cells in laminin-rich extracellular matrices (three dimensional or 3D culture) offers significant advantages over that in the conventional two-dimensional (2D) tissue culture system in that it takes into considetation the impact of extracellular matrix (ECM) microenvironment on the proliferation, survival, and differentiation of mammary epithelial cells. When grown in the 3D culture, untransformed mammary epithelial cells undergo morphogenesis to form a multicellular and polarized acini-like structure that functionally mimics the differentiated alveoli in the pregnancy mammary gland. This process is subjected to regulation by many growth factors and cytokines. The transforming growth factor-ß (TGFß) is a multipotent cytokine that regulates multiple aspects of development and tumorigenesis. In addition to its effects on epithelial cell proliferation, survival, and differentiation, it is also a potent regulator of the cell–matrix interaction. Thus, the 3D culture model may recapitulate the complex in vivo epithelial cell microenvironment and allow us to fully evaluate the role of TGFß signaling in multiple aspects of normal and cancerous cell behavior. In this chapter we provide detailed protocols for growing mammary epithelial cells in the 3D Matrigel for analysis of signaling pathways.
体外乳腺发育的重构:C-MET和C-ERBB2信号传导对分支和肺泡形态发生的需求。
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