Isolation, Culture And Identification Of Mouse Tail Vascular Endothelial Cells

Vascular endothelial cells(VEC) are made up of a layer of flat epithelial cells, which attached to the inner surface of blood vessels, forming an inner wall of the blood vessel. Vascular endothelial cells have a variety of functions, besides barrier function, it can also synthesize, secrete a variety of factors which play an important role in inflammation, injury, and immune response. Since the human vascular endothelial cell culture methods were reported, the external sources of vascular endothelial cells have been reported in recent years. The successful culture of endothelial cells in vitro, providing not only the study of vascular endothelial cells themselves, but also a good research tool for pathological and physiological effects of many diseases. At present, the culture methods of endothelial cells in vitro have tissue adherence, enzyme digestion and other methods, but each method has its own drawbacks, which needs further improvement and perfection.Angiostatin is one of the inhibitory factors, it can effective inhibit angiogenesis found in recent years. It can significantly inhibit the proliferation and migration of vascular endothelial cells and play an important role in the process of tumor metastasis. Therefore,Angiostatin can be used to inhibit the growth and metastasis of tumor cells. it is very promising in tumor treatment process, which is currently used in clinical trials.Hypericum perforatum Linn is belongs to Guttiferae. Hypericum extracts have a variety of chemical composition. It has proved of the therapeutic effects in depression, tumor and virus.Thus, after culturing vascular endothelial cells in vitro, we have further observed the biological activity of angiostatin and Hypericum perforatum Linn extracts on the cell proliferation.Main results and conclusions: 1. The culture of vascular endothelial cells from mouse tail blood vesselAfter stripping the blood vessels from mouse tail, carefully remove the adhesive tissues on the surface of the blood vessels, vascular surface were digested with collagenase. After cutting the blood vessels into small pieces, move the pieces into cell culture plate; cove the pieces with matrix gel and then adding culture medium. After three days of culture, the endothelial cells growth from the vascular edges, and gradually formed cell colonies. Under microscope, after the cell growth in confluence, the cells showed typical "cobble stone morphology. 2. Identification of vascular endothelial cellsThe identification of endothelial cell marker CD31 antibody showed more than 95% of the cultured cells are CD31 positive vascular endothelial cells, the result confirms the feasibility of the use of mouse tail blood vessels as the source of cultured vascular endothelial cells. 3.The biological activity of angiostatin on vascular endothelial cellsOn the basis of the vascular endothelial cells culture model, the biological activities of recombinant human angiostatin(rh K1-3) were tested on the Primary cultured vascular endothelial cells by MTT assay, it showed that the growth of cultured vascular endothelial cells were significantly inhibited by rh K1-3 in a dose-response manner. 3.The biological activity of Hypericum perforatum Linn on vascular endothelial cellsVascular endothelial cells were incubated in culture medium containing 0.025μg/ml、0.25μg/ml、2.5μg/ml Hypericum perforatum Linn by cell viability measurement by using MTT. 0.25μg/ml Hypericum perforatum Linn could significantly promote the survival of Vascular endothelial cells.