Platelet gel-released supernatant modulates the angiogenic capability of human endothelial cells

Hospital of L’Aquila, using multiplebags. Platelet gel was obtained by adding thrombin and calcium gluconate to the concentratesand then centrifuging to recover the supernatant.Human umbilical vein endothelial cells were isolated from umbil ical cord veins and grownin appropriate conditions. To study their viability, cells were treated with different concentrationsof supernatant and XTT assays were performed on the 3 days foll owing treatment. Endothelialcell motility and invasiveness were assayed using modified Boyd en chambers with filters coatedwith 0.1% gelatin (for the motility test) or with a thick layer of the reconstituted basementmembrane Matrigel (for the invasion test). The supernatant, add ed at various concentrations tothe lower compartment of the chamber, was used as an attractant . Umbilical cells were added tothe upper compartment of the chamber. After 4 hours (for the motility test) or 6 hours (for theinvasion test), filters were stained and the migrated cells in five high-power fields were counted.