2007Journal of Jilin UniversityRequires access

Effects of orientation transplantation of human bone marrow mesenchymal stem cells on behavioral tests of focal cerebral ischemia in rats

Nan Li

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Abstract

Objective To explore the effect of mesenchymal stem cells(MSCs) on nervous function in rats with focal cerebral ischemia.Methods The MSCs were cultivated,purified,and proliferated in vitro by density gradient and adherence to plastic dishes method.The models of Wistar rats were prepared after middle cerebral artery occlusion(MCAO) of right lasted 90 min and reperfusion 1 h.Wistar rats were randomly divided into normal control group(A,n=10),sham operation group(B,n=10),no-handle group after cerebral ischemia/reperfusion (C,n=10),free-serm DMEM transplantation group after cerebral ischemia/reperfusion(D,n=10),MSCs transplantation group after cerebral ischemia/reperfusion(E,n=10).After identified by flow cytometry,5 μL 5-bromo-2-deoxyuridine(BrdU) labeled MSCs(4×105· μL-1) and 5 μL serum-free DMEM were respectively injected intracerebraly into ischemic boundary zone of right in D and E groups.Immunohistochemical method was used to detect the expression and survival of BrdU-labeled MSCs in vivo.Nervous function behavioral tests were performed on 1st,3th,7th and 28th day after transplantation by forelimb use asymmetry test and postural reflex test.Results MSCs were successfully purified and proliferated in vitro.The MSCs expressed CD29,CD44,but didn't expressed CD34,CD45,CD31 identified by flow cytometry.transplanted MSCs survived and were localized to the ischemic boundary zone.Behavioral tests of every group were improved with time prolonged.However,MSCs transplantation group was significantly better than any other groups(P0.05).And on 7th day after MSCs transplantation,nervous function was significantly improved than other time points in MSCs transplantation group(P0.01).Conclusion The intracerebral administration of MSCs into ischemic boundary zone can promote the recovery of nervous function of rats with focal cerebral ischemia.Forelimb use asymmetry test can objectively evaluate motor function for a long time.

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Objective To explore the effect of mesenchymal stem cells(MSCs) on nervous function in rats with focal cerebral ischemia.Methods The MSCs were cultivated,purified,and proliferated in vitro by density gradient and adherence to plastic dishes method.The models of Wistar rats were prepared after middle cerebral artery occlusion(MCAO) of right lasted 90 min and reperfusion 1 h.Wistar rats were randomly divided into normal control group(A,n=10),sham operation group(B,n=10),no-handle group after cerebral ischemia/reperfusion (C,n=10),free-serm DMEM transplantation group after cerebral ischemia/reperfusion(D,n=10),MSCs transplantation group after cerebral ischemia/reperfusion(E,n=10).After identified by flow cytometry,5 μL 5-bromo-2-deoxyuridine(BrdU) labeled MSCs(4×105· μL-1) and 5 μL serum-free DMEM were respectively injected intracerebraly into ischemic boundary zone of right in D and E groups.Immunohistochemical method was used to detect the expression and survival of BrdU-labeled MSCs in vivo.Nervous function behavioral tests were performed on 1st,3th,7th and 28th day after transplantation by forelimb use asymmetry test and postural reflex test.Results MSCs were successfully purified and proliferated in vitro.The MSCs expressed CD29,CD44,but didn't expressed CD34,CD45,CD31 identified by flow cytometry.transplanted MSCs survived and were localized to the ischemic boundary zone.Behavioral tests of every group were improved with time prolonged.However,MSCs transplantation group was significantly better than any other groups(P0.05).And on 7th day after MSCs transplantation,nervous function was significantly improved than other time points in MSCs transplantation group(P0.01).Conclusion The intracerebral administration of MSCs into ischemic boundary zone can promote the recovery of nervous function of rats with focal cerebral ischemia.Forelimb use asymmetry test can objectively evaluate motor function for a long time.

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Available abstract

Objective To explore the effect of mesenchymal stem cells(MSCs) on nervous function in rats with focal cerebral ischemia.Methods The MSCs were cultivated,purified,and proliferated in vitro by density gradient and adherence to plastic dishes method.The models of Wistar rats were prepared after middle cerebral artery occlusion(MCAO) of right lasted 90 min and reperfusion 1 h.Wistar rats were randomly divided into normal control group(A,n=10),sham operation group(B,n=10),no-handle group after cerebral ischemia/reperfusion (C,n=10),free-serm DMEM transplantation group after cerebral ischemia/reperfusion(D,n=10),MSCs transplantation group after cerebral ischemia/reperfusion(E,n=10).After identified by flow cytometry,5 μL 5-bromo-2-deoxyuridine(BrdU) labeled MSCs(4×105· μL-1) and 5 μL serum-free DMEM were respectively injected intracerebraly into ischemic boundary zone of right in D and E groups.Immunohistochemical method was used to detect the expression and survival of BrdU-labeled MSCs in vivo.Nervous function behavioral tests were performed on 1st,3th,7th and 28th day after transplantation by forelimb use asymmetry test and postural reflex test.Results MSCs were successfully purified and proliferated in vitro.The MSCs expressed CD29,CD44,but didn't expressed CD34,CD45,CD31 identified by flow cytometry.transplanted MSCs survived and were localized to the ischemic boundary zone.Behavioral tests of every group were improved with time prolonged.However,MSCs transplantation group was significantly better than any other groups(P0.05).And on 7th day after MSCs transplantation,nervous function was significantly improved than other time points in MSCs transplantation group(P0.01).Conclusion The intracerebral administration of MSCs into ischemic boundary zone can promote the recovery of nervous function of rats with focal cerebral ischemia.Forelimb use asymmetry test can objectively evaluate motor function for a long time.

Key concepts: Transplantation, Mesenchymal stem cell, Ischemia, In vivo, Medicine, Flow cytometry, Pathology, CD34

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