Pellet-Many Caroline
Centre for Cardiovascular Biology and Medicine, University College London, Rayne Building, 5 University Street, London, WC1E 6JF, UK,
Methods Mol Biol. 2015;1332:151-7. doi: 10.1007/978-1-4939-2917-7_11.
In vitro assays of endothelial cell migration have led to critical insights into the mechanisms of angiogenesis. The transwell assay, or modified Boyden chamber assay was developed to investigate chemotaxis, which corresponds to the directional migration of cells in response to a chemoattractant gradient. It is a robust and easy-to-use assay that does not require expensive equipment. In the modified Boyden chamber assay, two compartments are separated with a porous membrane through which cells can migrate. The lower compartment contains the chemoattractant, creating a gradient by diffusing into the upper chamber containing the cells. The cells will migrate through the membrane and remain on the lower side of the membrane, where they can finally be fixed and counted.
内皮细胞迁移的体外测定已为血管生成机制带来了关键见解。Transwell测定法,即改良的博伊登室测定法,是为研究趋化性而开发的,趋化性对应于细胞响应化学引诱剂梯度的定向迁移。这是一种强大且易于使用的测定法,不需要昂贵的设备。在改良的博伊登室测定法中,两个隔室由多孔膜隔开,细胞可通过该膜迁移。下隔室含有化学引诱剂,通过扩散到含有细胞的上隔室中形成梯度。细胞将穿过膜并留在膜的下侧,最终可在那里固定并计数。