Del Moral Pierre-Marie, Warburton David
Saban Research Institute, Childrens Hospital Los Angeles, Los Angeles, CA 90027, USA.
Methods Mol Biol. 2010;633:71-9. doi: 10.1007/978-1-59745-019-5_5.
Lung primordial specification as well as branching morphogenesis, and the formation of various pulmonary cell lineages, requires a specific interaction of the lung endoderm with its surrounding mesenchyme and mesothelium. Lung mesenchyme has been shown to be the source of inductive signals for lung branching morphogenesis. Epithelial-mesenchymal-mesothelial interactions are also critical to embryonic lung morphogenesis. Early embryonic lung organ culture is a very useful system to study epithelial-mesenchymal interactions. Both epithelial and mesenchymal morphogenesis proceed under specific conditions that can be readily manipulated in this system (in the absence of maternal influence and blood flow). More importantly this technique can be readily done in a serumless, chemically defined culture media. Gain and loss of function can be achieved using expressed proteins, recombinant viral vectors, and/or analysis of transgenic mouse strains, antisense RNA, as well as RNA interference gene knockdown. Additionally, to further study epithelial-mesenchymal interactions, the relative roles of epithelium versus mesenchyme signaling can also be determined using tissue recombination (e.g., epithelial and mesenchymal separation) and microbead studies.
肺原基的特化、分支形态发生以及各种肺细胞谱系的形成，需要肺内胚层与其周围的间充质和间皮进行特定的相互作用。肺间充质已被证明是肺分支形态发生诱导信号的来源。上皮-间充质-间皮相互作用对胚胎肺形态发生也至关重要。早期胚胎肺器官培养是研究上皮-间充质相互作用的非常有用的系统。上皮和间充质形态发生均在特定条件下进行，这些条件在该系统中（在没有母体影响和血流的情况下）很容易操控。更重要的是，这项技术可以在无血清、化学成分明确的培养基中轻松完成。使用表达蛋白、重组病毒载体和/或分析转基因小鼠品系、反义RNA以及RNA干扰基因敲低，可实现功能的获得和丧失。此外，为了进一步研究上皮-间充质相互作用，还可以使用组织重组（如上皮和间充质分离）和微珠研究来确定上皮与间充质信号传导的相对作用。
