3.1.1 Boyden Chamber Assay for Endothelial Cell Migration
The Boyden chamber is a widely used experiment for evaluating the endothelial cells (EC) movement in response to chemotaxis. A polycarbonate filter with a particle size of 5-12 m separates the two compartments(Bahramsoltani et al., 2009). Extracellular matrix (ECM) protein, such as gelatin, collagen, fibronectin, or complex matrices like matrigel, can be coated on the filter. The upper chamber is seeded with ECs, while the lower chamber is supplied with culture media containing pro- or anti-angiogenic agents. The coatings can include anti-angiogenic substances. A layer of cancer cells beneath the filter can be used as a non-contact experiment to create a gradient of angiogenic agents (Van Moorst and Dass, 2011). The Image processing of labelled cell nuclei makes it simple to analyse EC migration via the filter(Iwahana et al., 1996).Enhanced sensitivity, high repeatability, and quick duration are all advantages of this assay (4-6 hrs)(Staton et al., 2009). The difficulty in observing cell vertical movement through a ‘foreign’ matrix is one of the disadvantages (polycarbonate or polypropylene filter)(Staton et al., 2009).