Spheroid-based human endothelial cell microvessel formation in vivo
Spheroid-based human endothelial cell microvessel formation in vivo
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DOI：
10.1038/nprot.2009.96
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发表时间：
2009-01-01
期刊：
影响因子：
14.8
通讯作者：
Augustin, Hellmut G.
中科院分区：
文献类型：
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作者：
Laib, Anna M.;Bartol, Arne;Augustin, Hellmut G.
近年来，血管生成内皮细胞（ECs）的研究极大地刺激了血管生物学研究的多个领域。许多细胞模型和许多复杂的发育，操纵和肿瘤动物模型已被开发用于研究血管生成在体外和体内。为了将细胞测定的多功能性与体内实验的读数的复杂性连接起来，我们开发了内皮移植测定。该测定基于在免疫受损小鼠中将离体产生的EC球状体（2d）移植到合适的基质中，以产生毛细血管的3D网络（20d）。该脉管系统连接到小鼠脉管系统，通过募集小鼠壁细胞来灌注和成熟。在这里，我们描述了详细的协议，这种测定，包括生成球状体，注射到小鼠，切除和处理所得的插头，并通过免疫组织化学分析的定量所得的脉管系统。
The study of angiogenic endothelial cells (ECs) has in recent years greatly stimulated multiple fields of vascular biology research. A number of cellular models and numerous complex developmental, manipulatory and tumor animal models have been developed to study angiogenesis in vitro and in vivo. To connect the versatility of cellular assays with the complexity of readouts of in vivo experimentation, we have developed an endothelial transplantation assay. This assay is based on grafting ex vivo generated EC spheroids (2 d) in a suitable matrix in immunocompromised mice, to give rise to a 3D network of capillaries (20 d). This vasculature connects to the mouse vasculature, is perfused and matures by recruiting mouse mural cells. Here we describe the detailed protocol for this assay, including generation of spheroids, injection into mice, excision and processing of resulting plugs, and quantification by immunohistochemical analysis of the resulting vasculature.