Neural Stem Cells Modified By Cytosinedeaminase Gene Control C6 Cells To Proliferate

Objective:We established a experiment that Neural stem cells modified by cytosine deaminase gene control C6 cells to proliferate.In order to find a new method to treat patient in glioma.Materials and Methods:1. Wistar foetus rat cerebral cortex and subcortex tissues were dissociated mechanically and enzymatically. The suspension culture technique was performed to acquire cell clones. the method of finity dilute was performed to acquqre inferior cell clones root in same cell.And immunofluorescene was used to detect nestin antigen of the clone2.CD gene was amplified and the pCMVCD plasmid was used to transfect the NSCs in vitro using lipofectamine2000 cationic lipid reagent.after gene transfection,the cells were screened by G418 presence,to detect immunal stably express NSCs were obtained3.NSCs/CD co-culture with C6 cells,separate group to observe Neural stem cells modified by cytosine deaminase gene control C6 cells to proliferate.to detect with MTT Data were expressed as x±s statistical analysis was performed with one-way ANOVA which was provided by spss 10.0 software significance was accepted at the level of p<0.05 and p<0.01 is very significance.Results:1.In this experiment ,we successful To establish the methods of harvest and culture of NSCs and to observe the characteristics of proliferation and induce differentiation of NSCs.Immunocytochemical staining was used to identify the NSCs. We have found that cell clones could be acquired from Wistar foetus rat cerebral cortex and subcortex tissues after primary and passage culture. The differentiatied cells express neurons,astrocytes and oligodendrocytes specific characteristics.2.pCMVCD plasmids were transfected by Lipofectamine 2000 liposomes conduct successful into NSCs .and the cells were screened by G418 presence,There clones were as a result of nestin-positive in immunofluorescence double mark,shew green fluorescence and at the same time expressed CD gene,shew red fluorescence.3.In the co-culture research ,we have finded Neural stem cells modified by cytosine deaminase gene control C6 cells to proliferate with 5-FC(100μg/mL).In the proportion 1:1,1:4,1:8 of the NSCs/CD and C6 cells,the cells which it grow with sticking to wall were patency reduce after three days.In the proportion 1:16,1:32,1:64,the cells have been controled too.the cells livability were in the difference 4.8%,5.2%,5.7%,18%,42%,84% after MTT check.Conclusion:The by-stander which Neural stem cells modified by cytosine deaminase gene control C6 cells to proliferate is very effective.It is owe to transferring ability of neural stem cells,It transfer around the glioma cells to send cytotoxin to it to die,these cells is good tool to send cytotoxin to glioma cells,the farther is very worth researching.