Koh Wonshill, Stratman Amber N, Sacharidou Anastasia, Davis George E
Department of Medical Pharmacology and Physiology, School of Medicine, Dalton Cardiovascular Center, University of Missouri-Columbia, Columbia, Missouri, USA.
Methods Enzymol. 2008;443:83-101. doi: 10.1016/S0076-6879(08)02005-3.
Discovery and comprehension of detailed molecular signaling pathways underlying endothelial vascular morphogenic events including endothelial lumen formation are key steps in understanding their roles during embryonic development, as well as during various disease states. Studies that used in vitro three-dimensional (3D) matrix endothelial cell morphogenic assay models, in conjunction with in vivo studies, have been essential to identifying molecules and explaining their related signaling pathways that regulate endothelial cell morphogenesis. We present methods to study molecular mechanisms controlling EC lumen formation in 3D collagen matrices. In vitro models representing vasculogenesis and angiogenesis, whereby EC lumen formation and tube morphogenesis readily occur, are described. We also detail different methods of gene manipulation in ECs and their application to analyze critical signaling events regulating EC lumen formation.
发现并理解包括内皮管腔形成在内的内皮血管形态发生事件背后的详细分子信号通路，是了解它们在胚胎发育以及各种疾病状态中所起作用的关键步骤。将体外三维（3D）基质内皮细胞形态发生分析模型与体内研究相结合的研究，对于识别分子并解释其调节内皮细胞形态发生的相关信号通路至关重要。我们展示了研究三维胶原基质中控制内皮细胞管腔形成分子机制的方法。描述了代表血管生成和血管新生的体外模型，在内皮细胞管腔形成和管形态发生很容易发生的情况下进行了描述。我们还详细介绍了内皮细胞中基因操作的不同方法及其在分析调节内皮细胞管腔形成的关键信号事件中的应用。
