Thestudy Of Culturing Swine Umbilical Veinendothelial Cells In Vitro

Objective:To provide a good cell model for the study about the impact o f the swin eplacenta and placental angiogenesis which the Peroxisome prolifer ator-activated receptor gama gives, the experiment cultures the swine umbilical vein endothelial cells in vitro and make them immortalized.Methods:Use collagenase to digest and separate fresh swine umbilical vei n endothelial cells, and culture them in vitro by the CO2 Incubator;observe the morphological changes of cell growth through microscope and make them ide ntified by immunohistochemical method; monitor the cell growth curve by using iCELLigence cell function analyzer and the cell cycle and apoptosis are dete cted by fow cytometry;the experiment make the primary cells immortalized by making telomerase in,and then get cells for monoclone and amplification by u sing G418;the study about the effect of PPARy which regulate and control by its agonist rosiglitazone and inhibitors T0070907 on cell proliferation by using cell function analyzer.Results:(1)Primary cells who have clear boundaries can grow monolayer well which show small triangle? oval? Spindle and Polygon,and they make cob blestone-like arrangement after cellular fusion which have strong proliferation. B ut, the cells have weak cell proliferation and poor cell viability,increased apopt osis after several times of passaging.The results of cell identification are all posi tive by using F? Ag and CD31.(2) Telomerase prove successful into the cells after make it in though the detect of transfection efficiency;the experiment gains monoclonal cells after 14 days'drug Screening, and they can well grow and proliferate.(3)Both the primary cells and the monoclonal cells can grow fast,have str ong cell viability and the expression of PPARy protein can be increased when the agonist concentration is 5uMol/ml;the expression of PPAR? protein is inhibi ted significantly,cell growth and proliferation are also been inhibited,and cell vi ability is decreased when the inhibitor concentration is 20mMol/ml,no matter th ey are primary cells or monoclonal cells.Conclusion:The study culture the cells which separated from swine umbilical vein in vitro,and they are proved according with the basic characteristics of vascular endothelial cells through observing and cell characteristic markers detecting;the experiment gains monoclonal cells which can grow well and have good proliferation ability after telomerase transfection and drug Screening;both the primary cultured cells and the transfected monoclonal cells show the reaction of promoting or inhibiting cell growth.