Experimental Investigation Of The Effect On MSCs Transplantating Through Different Paths After Cerebral Ischemia In Rats

Objective:To observe the recovery of neurological function and investigate the protective mechanism and select correct transplanting path in rats following Cerebral Ischemia.After mesenchymal stem cells(MSCs)of Middle cerebral Artery Occlusion(MCAO)rats were transplanted by stereotaxis and vena caudalis path.Methods:Bone marrow cells were isolated,proliferated,purified and passaged from the long bone of 150g SD rats.MSCs were preinductived 24 hours by FGF-2.MSCs (preinductived and uninductived)were marked by Brdu(10mg/L)before 24 hours transplanted.Then were digested for animal experiment;MCAO model was established with improved Zea Longa suture emboli method.To judgment site and scope of cerebral ischemia with MRI on head;40 MCAO SD rats were devided into 5 groups,8 rats in each group.(Ⅰ,Ⅱ)Stereotaxis transplanted group and(Ⅲ,Ⅳ)vena caudalis transplanted group,(Ⅴ)control group at random,preinductived and uninductived groups were in cluded in transplanted groups.Ischemic model was made in model group only.MSCs were labeled by BrdU were injected into MCAO rats respectively through stereotaxis and vena caudalis paths 1 week after made modles,MSCs were injected into corpus striatum of ischemia injured side.Then①Compare NSS at different time(1d,2w,1m,2m,3m).②The rats were executed at 1m,2m and 3m.Their brain tissues were poly-methanal-fixed,paraffin-embed,and paraffin sections.③To evaluate the relationship between survivorship,migration and differentiation,Brdu,NSE and GFAP were choosed as the markers of proliferation and differentiation.Brdu+NSE,Brdu+GFAP and BDNF were identified by immunohistochemisty staining.④To investigate the repair's mechanism of MCAO rats by analyzing the conditions of MSCs' survivorship,migration,differentiation through different transplanted path.Results:MSCs got purified through isolation and passage,morphous of cells were Fusiform,whirlpoolly.By using FRCS,the masccline rate of CD90 is 99%,CD34 is 0.3%,CD31 is 3.4%,The isolated cells were MSCs;The opposite side limb paralysis after MCAO with a nylon suture.MRI showed succeed models were made.The peak time of NSS in each group is 1d after transplanted.There were no difference(p＞0.05);The NSS of the transpalnted groups were lower than that of contral group at 2w,1m,2m,3m(p＜0.05);The scores of NSS inⅢ,Ⅳgroup was lower than that ofⅠ,Ⅱgroup at 2w(P＜0.05).There were no difference in transplanted groups at 1m(p＞0.05);The stereotaxis group's were lower than vena caudalis group's at 2m,3m;The preinductived group's were lower than uninductived at 2m(p＜0.05);There were no difference of statisticst between preinductived group and preinductived group at 2w,1m,3m.Brdu+NSE and Brdu+GFAP can be seen in the stereotaxis group and the vena caudalis group at 1m by immunohistochemisty staining.There were more double positive cells in preinductived group than that's in preinductived group at 2m, however there were no obviously different at 3m.There were a lot of Brdu positive cells in transplanted area of ischemia hemisphere and distribute around pinhole in the strerotaxis group,wherever no in the opposite side.The were a little in both hemispheres in vena caudalis group.The level of BDNF in the transplanting groups were much more than the contral group,the stereotaxis group's were higher than the vena caudalis group's(p＜0.05).Conclusion:MSCs can be isolated,passaged and proliferated;MSCs transplanted by the stereotaxis and vena caudalis can survival and different into neuron like cells and neuroglia like cells,and can improve neurological function.The level of BDNF was much higher in the transplanted group's and can make recorvery of neurological function of MCAO rats;Stereotaxis group's therapeutic efficacy is more obvious than that's of vena caudalis group.