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).