| Objectives:After we established SD rats middle cerebral artery occlusion(MCAO)models with modified zealonga method,mesenchymal stem cells(MSCs)labeled with green gluorescent protein(GFP)were transplated into SD rats through directly stereotactic injection into brain ventricle,intracarotid administration and internal jugular vein infusion.The impact on neurological function after transplantation of bone mesenchymal stem cells(BMSCs)is observed.Whether the transplanted BMSCs have the ability of homing to the injured cerebral tissue and the distribution of the stem cells in cerebral tissue are also observed.How to pass through the blood-brain barrier and the possible mechanism of the transplanted BMSCs accelerating the repair of cerebral ischamic is discussed.We also compare the therapeutic efficacy of three ways of transplantion.Methods:①Resouce and culture of BMSCs:BMSCs labeled with GFP come from the cell line established by our laboratory.BMSCs were cultivated by adherent culture.②Characterization of the cultured cells:Study the Cellular morphology with a phase-contrast microscope and observe the living characteristics after subcukture of the cells.Identify the cells by immunohistochemiacal assessment.③Identify cerebral infarction region after MCAO by 2,3,5-triphenyl-tetrazolium chloride(TTC)staining.④Evaluation after tansplated BMSCs into rats:When the third day after established MCAO model,we tansplated BMSCs into rats through three ways above,which including directly stereotactic injection into brain ventricle,intracarotid administration and internal jugular vein infusion.Then we compare recovery of neurological function in three ways by 5-scoring system refer to Longa and Berdeson's methods in the exoerimental and contral groups.⑤Under fluorescence inverted microscope quantity of survival fluorescent cells and migration and distribution regularityrats in brain tissues slices were surveyed.Results:①Culture and characterization of BMSCs labeled with GFP:After 24 hours of culture,there are many rounded cells and some heterogeneous small spindle-shaped fibroblastoid cells growing in the culture flask.After 3 days of culture,BMSCs grew in colonies that more heterogeneous small spindle-shaped fibroblastoid cells and less rounded cells.After 4 days of culture,the amplified BMSCs were homogeneous, spindle and colony-shape.BMSCs were positive for CD29,CD44＞98%and negative for CD34.②5-scoring system about recovery of neurological function:There was no significant difference between rats with PBS and rats with BMSCs before transplantation.Rats transplanted with BMSCs had significantly lower scores at day 14 compared with the control group.③The amount of GFP positive cells within damaged areas in the transplantation group were larger than that in the symmertrical brain area and the other brain areas in the same group as in the same brain area in control groups.Conclusions:①BMSCs transplated into rats has the evident reparative effects on cerebral ischemic injury.②The transplanted BMSCs are capable of homing to injured cerebral tissue.They can accelerate the recovery of neurological function.The amount of GFP positive BMSCs in the damaged areas is maximum.③It is the best ways to transplate into rats through directly stereotactic injection into brain ventricle... |
