Aplin Alfred C, Fogel Eric, Zorzi Penelope, Nicosia Roberto F
Department of Pathology, University of Washington, Seattle, Washington, USA.
Methods Enzymol. 2008;443:119-36. doi: 10.1016/S0076-6879(08)02007-7.
Angiogenesis is regulated by a complex cascade of cellular and molecular events. The entire process can be reproduced in vitro by culturing rat or mouse aortic explants in three-dimensional biomatrices under chemically defined conditions. Angiogenesis in this system is driven by endogenous growth factors released by the aorta and its outgrowth. Sprouting endothelial cells closely interact with pericytes, macrophages, and fibroblasts in an orderly sequence of morphogenetic events that recapitulates all stages of angiogenesis. This model can be used to study the basic mechanisms of the angiogenic process and to test the efficacy of proangiogenic or antiangiogenic compounds. Aortic cultures can be evaluated with a range of morphologic and molecular techniques for the study of gene expression. In this chapter we describe basic protocols currently used in our laboratory to prepare, quantify, and analyze this assay.
血管生成受一系列复杂的细胞和分子事件调控。通过在化学限定条件下将大鼠或小鼠主动脉外植体培养于三维生物基质中，整个过程可在体外重现。该系统中的血管生成由主动脉及其生长物释放的内源性生长因子驱动。发芽的内皮细胞与周细胞、巨噬细胞和成纤维细胞以有序的形态发生事件序列紧密相互作用，概括了血管生成的所有阶段。该模型可用于研究血管生成过程的基本机制，并测试促血管生成或抗血管生成化合物的功效。主动脉培养物可通过一系列形态学和分子技术进行评估，以研究基因表达。在本章中，我们描述了目前在我们实验室中用于制备、定量和分析该测定的基本方案。
