In Vitro Tube Formation Assay

Angiogenesis, or the formation of new blood vessels, plays an important role in many physiological (e.g. wound healing) and pathological (e.g. tumor growth and inflammation disease) processes. As a complex multi-step process, angiogenesis involves degradation of basement membrane, proliferation, and migration of endothelial cells into extracellular matrix (ECM), followed by alignment and reorganization of endothelial cells into three-dimensional tubular structure and formation of new basement membrane. Specifically, the critical step of the formation of three-dimensional tubular structure by endothelial cells is the objective of many in vivo and in vitro angiogenesis assays to screen efficacy of both pro- and antiangiogenic agents. The ScienCell(TM) In Vitro Tube Formation Assay (IVTFA) provides a convenient system to measure the formation of three-dimensional tubular structure by endothelial cells when cultured on an ECM gel of basement membrane extract. The ECM gel, which is prepared from Engelbreth Holm-Swarm (EHS) sarcoma produced in mice, consists of laminin, collagen IV, heparan sulfate proteoglycan, entactin and other minor components optimized for tube formation. With this system, tube formation can be achieved within 6-18 hours, with the earliest signs of tubular structure shown up after 3-4 hours. The Assay also provides a colorimetric cell staining reagent for better examination.