Treatment Of Ischemic Stroke: The Synergy Between Microglia Elimination And Neural Stem Cell Transplantation

The neural stem cells from neonatal mice cultivation and identification in vitroOBJECTIVE: Established isolation culture system of neural stem cells from newborn mice hippocampus and identif ication the product.Methods: The serum-free suspension culture technique was used to isolate and pass on the neural stem cells of the newborn C57 BL / 6 mice.The third generation neural stem cells treatment by Brd U and Ed U,nestin to identify neural stem cells,10% fetal bovine serum induced neural stem cells.Immunofluorescence staining with NS E and GFAP.Brdu/Hochest33342 and Edu/Hochest33342 double-labeled positive cells were counted and statistical analysis,comparing the tagging efficiency of Brd U and Ed U.Results: The hippocampus-derived neural stem cells of neonatal mice could be cultured in vitro,and the results of nestin staining were positive.In vitro,serum-induced differentiation could produce specific enolase and glial fibr illary acidic protein positive cells.The group of Brd U and Ed U have a signif icant(p<0.05).Conclusion: The neural stem cells obtained by in vitro isolation and culture were used to reproduce the multiplication and multiplication ability of the neural stem cells.Ed U was better in tagging efficiency and more sensitive.The neural stem cells of neonatal mice hippocampus cryopreservation and recoveryOBJECTIVE: To observe the survival rate and proliferation ability of neural stem cells after cryopreservation with different cryopreservation solutions and time.Methods: Taking two kinds of solution cryopreservation neural stem cells then after 1 months,3 months and 6 months of recovery.Trypan blue was used to detect the percentage of viable cells after resuscitation then CCK-8 detecting the proliferation capacity of neural stem cells from 1 to 7 day.Results: The same kind of frozen liquid frozen cryopreservation neural stem cells at different time there was no signif icant of cell survival rate(p>0.05),at the same time using two frozen liquid cryopreservation cells after recovery of the cell survival rate was statistically signif icant(p<0.05).CCK-8 was used to detect the proliferation ability of the cells with different solution and cryopreservation time,there was statistically s ignificant between the groups at sixth days(p<0.05).Conclusion: The two kinds of solution can be used in the cryopreservation of neural stem cells,with the cryopreservation time extension proliferation ability of neural stem cells would be decreased.The research of neural stem cells transplantation for the treatment of ischemic strok eObjective: To observe the repair effect of NSCs transplant after acute ischemic stroke in mice model,explor ing the potential mechanism.Methods: The model of acute ischemic stroke was induced by photochemical method.1day before surgery and 1d,3d,7d,14 d,21d,28 d,35d post transplantation rotary test and grip strength test were conducted to evaluate behavioral outcomes,observed NSCs differentiated into neurons and astrocytes by immunofluorescence staining.Results: Rotary test: the group of B and C have no significant(p>0.05),group of B and D after transplantation of 1d have no significant(p>0.05),3d after transplantation(?p< 0.05),the other time points(?p<0.01).Grip strength test at each time point suggest that compare with B and C no signif icant(p>0.05),group of B and D 1d after transplantation no signif icant(p>0.05),the other time points there were significant(?p<0.01).Immunofluorescence staining results showed that Edu/GFAP and Edu/NG-2 double positive cells were observed at 5d after transplantation.Edu/GFAP and Edu/MAP-2 double positive cells were observed at 14 d after transplantation.After 28 d and 35 d transplantation,Edu/GFAP,Edu/MAP-2,Edu/Neu N double positive cells were observed.Conclusion: Exogenous NSCs transplantation can differentiated into neurons and astrocytes in vivo and contribute to neurogenesis recovery of behavioral function after ischemic stroke.Treatment of ischemic strok e: the synergy between microglia elimination and neuralstem cell transplantationObjective: Compare the therapeutic effect of transplantation of NSCs and microglial elimination synergistic NSCs transplantation in ischemic stroke,exploring the potential mechanism.Methods: Groups of A and B all accept exogenous neural stem cell transplantation,while in the B group after surgery beginning chow PLX3397 consecutive 7days,two groups were conducted rotary test,grip strength test and open field test.Ed U,Nestin Immunofluorescence staining at 3d after transplantation.Ed U,GFAP,MAP-2 staining was used to observe the NSCs differentiation.Results: Rotary test: groups of A and B have signif icant at 7d,14 d,21d,28 d,35d after transplantation(?p<0.05).Grip strength test showed that at 14 d,21d,28 d,35d after transplantation there was significant between A and B(?p < 0.05;?p<0.05),at 35 d after transplantation there was no significant(p>0.05).Immunofluorescence staining showed that B group was better than group A.Conclusion: Microglial elimination synergistic NSCs transplantation is more beneficial to the survival and differentiation of the NSCs.