Watanabe Motomu, Fujioka-Kaneko Yayoi, Kobayashi Hisayuki, Kiniwa Mamoru, Kuwano Michihiko, Basaki Yuji
Advanced Research Laboratory, Taiho Pharmaceutical Co., Ltd, Saitama, 357-8527, Japan.
Biol Proced Online. 2005;7:41-7. doi: 10.1251/bpo104. Epub 2005 Apr 27.
Angiogenesis is a complex process involving an ECM and vascular endothelial cells (EC), and is regulated by various angiogenic factors including VEGF. The ability to form a capillary/tube-like network is a specialized function of EC. Therefore, in vitro angiogenesis was assessed by a capillary/tube-like network formation assay. There are three angiogenic parameters: capillary length, number of capillaries, and relative capillary area per field. We evaluated capillary length per field in the assay. VEGF promoted capillary/tube-like network formation of EC in a type I collagen gel matrix in vitro. Moreover, we demonstrated the involvement of ILK in a VEGF signaling pathway mediating capillary/tube-like network formation of EC using dominant-negative, kinase deficient ILK. This is a straightforward assay to monitor responses of human vascular endothelial cells.
血管生成是一个涉及细胞外基质(ECM)和血管内皮细胞(EC)的复杂过程,并受包括血管内皮生长因子(VEGF)在内的多种血管生成因子调节。形成毛细血管样或管状网络的能力是内皮细胞的一项特殊功能。因此,通过毛细血管样或管状网络形成试验评估体外血管生成。有三个血管生成参数:毛细血管长度、毛细血管数量以及每视野的相对毛细血管面积。我们在试验中评估了每视野的毛细血管长度。VEGF在体外I型胶原凝胶基质中促进内皮细胞的毛细血管样或管状网络形成。此外,我们使用显性负性、激酶缺陷型整合素连接激酶(ILK)证明了ILK参与介导内皮细胞毛细血管样或管状网络形成的VEGF信号通路。这是一种监测人血管内皮细胞反应的直接试验。