Magnetic bead assay for mechanical stimulation of endothelial cells

The assay mechanically stimulates endothelial cells using thrombin-coated magnetic beads that are manipulated by rare earth magnets. It can more specifically study the effects of mechanical stimulation compared to using soluble signaling factors. Moreover, somewhat variable static and dynamic stimulation studies can be implemented by adjusting the placement and rotation speeds of the magnets.

BD Horizon™ BV510 Mouse Anti-Human CD45RA Antibody
RRID:AB_2722499

BD Pharmingen™ PE-Cy™5 Mouse Anti-Human CD56 (NCAM-1) Antibody
RRID:AB_395907

Vasculogenic ring assay

The assay concept is embedding a 1:1 ratio of endothelial cells and fibroblasts in a fibrin gel to study vessel formation in vitro and in 3D. Fibrin density of the gel can be varied. After 7 days, vessel growth (ring) can be analyzed and imaged by staining the hydrogels for the endothelial marker CD31. Soluble VEGF produced during the ring assay can be measured using VEGF Human ELISA Kit. VEGF signaling can be inhibited by adding a small molecule inhibitor of VEGFR, SU-5402.

PE anti-human TCR γ/δ Antibody
RRID:AB_1089219

CD8b Antibody
RRID:AB_2659521

PE anti-human CD107a (LAMP-1) Antibody
RRID:AB_1186040

Microfluidic device to attain high spatial and temporal control of oxygen
RRID:SCR_017131
The microfluidic device can control both the spatial and temporal variations in oxygen tensions to better replicate in vivo biology. The device consists of three parallel connected tissue chambers and an oxygen scavenger channel placed adjacent to these tissue chambers. Experimentally measured oxygen maps were constructed using phosphorescent lifetime imaging microscopy and compared with values from a computational model. For testing, it was coupled with a 3D sprouting angiogenesis assay. The simple design provides consistent control of spatial and temporal oxygen gradients in the tissue microenvironment and can be used to investigate important oxygen-dependent biological processes.