Experiment Study Of Cultivation And Characteristics Of Schwann Cell In Vitro

Objective: To present an effective technique for isolation , cultivation and purification of Schwann cell (SC) from newborn(2-5 days) rat peripheral nerves .And to observe the growth rule of SC in Vitro and the effect of bFGF(basic Fibroblast Growth Factor) on it .Methods : Newborn (aged 2-5 days) SD rat sciatic nerves were harvested .After the sciatic nerve tissue pieces were digested by two enzymes for few minutes ,purified SC were obtained after the pieces were implanted repeatedly and Geneticin was added in the cultured medium in order to inhibit fibroblasts . The cultured cell was stained by S-100 with immunochemical methods .The propertied of proliferation of SC in Vitro was observed ,and the growth curve of SC 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 bFGF on SC with MTT method and flow cytometry .Results : Most of fibroblasts were killed and purified SC were obtained .S-100 stain showed that the purity of SC population was up to 95% .After sub-culture, the purity of SC population was up to 98% .The quantity of SC was up to 1×107/ml .The growth curve of SC was drew , and the doubling time of the 2nd passage of cells was 8 days .The density of SC in 1ng/ml bFGF group and 10ng/ml bFGF group were more than that in non-bFGF group ,the significant difference between them was detected (P<0.001) with the MTT method .The number of SC in S phase and G2M phase in 10ng/ml bFGF group was more than that in non-bFGF group ,the significant difference between them was detected (P<0.01) with the flow cytometry .Conclusion: Plenty and purified SC are obtained with these methods .bFGF in proper concentration could promote the proliferation of SC .These findings provide a source of SC for tissue engineering cellular transplants.