Experimental Study Of Cultivation,Generation Characteristics And Stimulus Response To Anti-TGF-β Of Fibroblast From Peripheral Nerve Impaired In Vitro

Objective: To present an effective technique for isolation , cultivation and purification of Fibroblast from(aged 30 days)Wistar rat Peripheral Nerve impaired .And to observe the proliferation characteristic of Fibroblast in Vitro and the effect of anti-TGF-β(Transforming Growth Factorβ) on it .Methods : Wistar rat (aged 30 days) sciatic nerves were cut off .Under the microscope,epineurium is sewed up . After breeding 12-15 days, sciatic nerves in animal model were harvested.Adopt the tissue where the nerves were cut off . The tissue snipped into pieces(1mm3) was cultivated for a few time . Through a series of purification methods , purified Fibroblast were obtained. . The cultured cell was stained by vimentin with immunochemical methods .The propertied of proliferation of Fibroblast in Vitro was observed ,and the growth curve of Fibroblast was drew .The doubling time of the 2nd passage of cells was worked out by the formula for the doubling time of the cell . To study the effect of anti-TGF-βon Fibroblast with MTT method . To determine theⅠtype collogen from Fibroblast with enzyme linked immunosorbent assay(ELISA).Results :Through the method used in the study ,a lot of Fibroblast were obtained. Its purity was up to 98%. Vimentin stain showed that it was what we need. The growth curve of Fibroblast was drew, and the doubling time of the third or the forth passage of cells was 3 days . In comparison with the no-anti-TGF-βgroup ,the two groups (10ng/ml, 100ng/ml) get more the density of Fibroblast . The significant difference between them was detected with the MTT method .( P<0.05 ) Different densities of anti-TGF-βwere used to inferere in the growth of Fibroblast , and then collect the cell supernantants . In the two groups (10ng/ml, 100ng/ml) , Fibroblast growth was obviously hold dack and the production ofⅠtype collogen was reduced . Comparion with the control , the significant difference was detected with the ELISA method .( P<0.05 )Conclusion: Using the method provided in the study , we can get more plenty and more purified Fibroblast from Peripheral Nerve impaired . Debita spissitudine anti-TGF-βcan inhibit the growth of Fibroblast and reduce the production ofⅠtype collogen . It is evidenced that anti-TGF-βhave depressant effect on nerve scar .