Study On Folic Acid Combined With Neural Stem Cells In The Treatment Of Focal Cerebral Ischemia Injury Brain In Rats

Part1:Isolation,culture and identification of neural stem cells from newborn mouse spinalObjective: To separate training new spinal cord neural stem cells in mice, proliferation, differentiation and identification, in order to further study the biological characteristics of spinal cord neural stem cells.Method: Taking 6 newborn mice, sterile precooling conditions in spinal cord tissue, using the mechanical separation blowing into a single cell, primitive cells using serum-free culture, the mechanical method of trypsin digestion method is adopted to develop and extend the primary cell. Application of immunofluorescence nestin, dyeing, CD133 tubulin(beta tubulin Ⅲ), glial acidic protein(GFAP) fiber dyeing, identification of spinal cord neural stem cells and differentiated products.Results: the neural stem cells from the mice in the spinal cord has the ability to clone cloned cell nests protein, CD133 positive immune reaction. Join to induce differentiation of fetal bovine serum to beta tubulin-Ⅲ and GFAP positive cells.Conclusion: mechanical separation method of new spinal cord cells in mice, mechanical composite enzyme digestion method to extend the primary cell and to develop, to success in the spinal cord neural stem cells, can be induced to differentiate into neurons and radial glial cells. Part2:Study on Folic acid combined with spinal neural stem cells in treatment of focal cerebral ischemia in miceObjective:Study on Folic acid combined with spinal neural stem cells in treatment of focal cerebral ischemia in mice and explore its possible mechanism.Method:sixty Rats were randomly assigned to 3 groups: MCAO; MCAO and exogenous NSC transplantation; and MCAO, NSC transplantation and FA.FA(0.8 mg/kg) or vehicle was administered by gavage daily for 28 days before MCAO and 23 days afterward.A week each group serum folate concentration is measured.Animal model of cerebral ischemia in mice is established by Photoinduced cortex thrombosisd.Neural stem cells is transplantated, which cerebral infartion is established.slices is dyed by Fluo-Jade staining and Nissl’s staining.eurons is counted.DDouble positive cells number is detected by immunofluorescence double labeling.The brain tissue expression of apoptosis gene Bax and Casosse-3 is by plague imprinting method.Results:Compared with group A and group B,group C serum folate concretrantion incressed significantly. Compared with group A,group C parietal cortex neuron numberincreased.Compared with group A,group C parietal cortex neuron number increase.Compared with group A,group C double positive cells number increase.Compared with group A,group C apoptosis protein Bax and Casosse-3 increase.Conclusion:By folic acid,serum folate concentration is improved.It can promote the suivival and differentiation of neural stem cells.It can Reduce the apoptosia of cells in the central nervous so toachieve therapeutic effect.