The Experimental Study On Culturing,identifying, SPIO Labeling And MRI Tracing In Vivo Of Rat Bone Marrow-derived Mesenchymal Stem Cells

ObiectiveIsolate,culture,identify and Superparamagnetic iron oxide(SPIO) lable of rat bone marrow-derived mesenchymal stem cells(BMSCs), to evaluate the feasibile value of tracing in vivo by 3.0 Tesla magnetic resonance imaging .Methods1.Isolate and culture rat BMSCs by attachment method. Identification was performed by flow cytometry ,osteoblasts and adipocytes differentiation,and study the influence of different modalities of the first medium exchange on culturing.2.BMSCs endocyte SPIO on the conditation of transfection agent PLL,and then evaluate the biologic characteristics and the optimal concentration of SPIO.3.Construct TBI model by reference of Feeney method, inject and transplant SPIO labled BMSCs trans-right common carotid artery.4.The experimental group rat were injected SPIO labled BMSCs,and the control group 1 were injected non-SPIO labled BMSCs, the control group 2 were injected pu- re PBS only.All 3 groups were analyzed by NSS and MR scaning at the point of pre- injecting and post-injectiong 1d,3d,7d,14d,21d,observe the signal exchange ang its migration in brain tissue,and compare with the contralateral signal of brain tissue. After every MR scaning,a rat was killed and its brain was taken out and performed HE and Prussian blue staining.Result1.BMSCs grew well,and the culture period an purity were aviliable.FCM test showed CD29,CD90 postive while CD34,CD45 negative.BMSCs were induced to differ- entate into osteoblasts and adipocytes,and show no influence on characteritics after labled by SPIO.2.In the experimental group,brain tissue emerged decreased signal after SPIO labled BMSCs injection,and decresed signal migrated towards to the lesion,which was significant different compared with the contralatera(lF=154.19,P<0.05).Blue particles were observed in brain sections afet Prussian blue staining.The BMSCs transplantation groups were lower NSS than PBS group(P<0.05).Conclusion1.Attachment method is suitable for BMSCs culturing.BMSCs can endocyte SPIO without biologic characteristics influencing.2.3.0T MRI shows value on tracing of SPIO-labled BMSCs in TBI modle of rat.3.BMSCs may play a role on the neural function recovery.