The Effects Of BMSCs Derived NSCs On Neurological Recovery After Spinal Cord Injury

Objective To create a methods by which rat's bone marrow mesenchymal stem cells(BMSCs)can be cultivated in vitro.To oberserve the possibility of differentiation of BMSCs into-neural stem cells(NSCs),then used in SCI,to explore the effects of BMSCsD-NSCs on neurological recovery after spinal cord injury.Methods Bone marrow were obtained from rats.Cells were primarily cultured by adherence method,and indentitied by the multipotential ability.The passaged ceils were resuspended and induced in the DMEM/F12 medium without serum containing EGF, bFGF,N2,B27.Immunocytochemistry was used to identify expression of nestin(maker for NSCs).And then we added serum,and immunocytochemistry was used to identify expression of MAP2(maker for neuron),GFAP(maker for astrocytes)and MBP(maker for oligo-dendrocytes).80 SD rats were randomized to four groups:normal(Group A,n=20), and injury(Group B,Group C and Group D;Given T₉ laminectomy with spinal cord left-hemisection,each group n=20);Group B was control group(given saline in injured spinal cord),Group C(given BMSCs-D-NSCs),and Group D(NSCs)were transplanted groups.Immunohistochemistry of BrdU were detected after injury.BBB were assessed by blinder observers at 1,2 and 4 weeks after surgery.Results The cultivation of BMSCs in vitro shows that they increased lastingly and has a fast multiplication speed in vitro.They could be passaged and amplified lots of generations,and could be differentiated into osteoblast cells,adipocyte and neuron-like cells.BMSCs in the medium containing EGF,bFGF,N2,B27 could differentiate into cellular spheres with positive nestin.Following differentiation of cellular spheres in the medium with serum,some of them formed neuron showed positive MAP2,some formed astrocytes showed positive GFAP,and others formed oligodendrocytes showed positive MBP and PLP.After the surgery,The BBB of group A was 21.Certain neurological recovery occur in Group B,C,D at 1,2,4 weeks after hemisection.BBB showed significant difference between Group B and Group C or D(P＜0.05),The BBB was no significant difference between Group C and D.7 d after transplanting in Group C and D,the cells labled BrdU could be observed in the spinal cord.And gradually the cells labled BrdU could't be observed after 28 d.Conclusion BMSCs could be differentiated into NSCs,and the BMSCs-D-NSCs have the ability to differentiate into neuron,astrocytes and oligodendrocytes.The neurologic function recovery were significantly improved in transplanted groups.