Endostatin Factor Thrombospondin-1 On Lymphatic Endothelial Cells Generated

Objective To study the effect and mechanism of thrombospondin-1 (TSP-1) on proliferation and migration of lymphatic endothelial cells (LECs) and further to find the safe, effective and practical inhibitors of lymphangiogenesis.Material and Methods LECs were taken from the thoracic duct of healthy pig. 1 Saparation,cultivation and passage of LECs. 2 Identifiction of LECs:The experiment adopt immunocyte chemistrity method,factor VIII related antigen and LEC peculiarity antibody vascular endothelial growth factor receptor-3(VEGFR-3) were used to identify LECs. 3 Inhibitory experiment:The experiment adopted method of scraping line and MTT to observe the proliferation and migration of LECs which were affected by thrombospondin-1. (1) Method of scraping line:It included control group and 3 experimental groups of different concentration. (2) MTT method:It included control group and 5 experimental groups of different concentration.Results 1 Identifiction of LECs:The cultivated cells were identified as typical LECs by factor VIII related antigen and VEGFR-3. 2 light microscope observation: under light microscope, LECs presented the typical character of"cobblestone"or"the flagstone". 3 Method of scraping line : Comparing with the control group,when concentration of thrombospondin-1 reached 1.0ug/ml,it could inhibit the proliferation and migration of LECs (P<0.01). 4 MTT method:When concentration of thrombospondin-1 reached 0.8ug/ml,it could significantly inhibit the proliferation of LECs(P<0.01).Conclusion 1 The cultivated cell which keep its biology character is the typical LEC. 2 The method which use the factor VIII and VEGFR-3 to identify the LECs is a special method.3 Thrombospondin-1 could evidently inhibit the proliferation and migration of LECs and were dose-dependent.