骨髓基质细胞体外分化移植治疗大鼠脊髓损伤的初步研究

Primary investigation of bone marrow stromal cells transplantation after differentiation on the spinal cord injury of rats

[目的]探讨大鼠骨髓基质细胞体外分化为神经干细胞后移植治疗大鼠脊髓损伤的可行性。[方法]骨髓基质细胞经培养及定向分化为神经干细胞,后者由5-溴脱氧尿嘧啶核苷法标记,制备大鼠脊髓损伤模型,伤后第9 d移植神经干细胞,实验分组:细胞移植组、PBS填充组、正常对照组。应用组化法观察移植细胞是否存活,取材前24 h显露坐骨神经,行辣根过氧化物酶逆行示踪法观察脊髓损伤处的修复重建。[结果]骨髓基质细胞在定向分化为神经干细胞后标记并移植于脊髓损伤区,标记的阳性细胞可在受体脊髓内检测到,辣根示踪技术显示细胞移植组较PBS填充组阳性细胞明显增多,差别有统计学意义。[结论]大鼠骨髓基质细胞在体外分化为神经干细胞后移植于脊髓损伤区,移植细胞可以存活,并参与脊髓损伤处神经传导通路的结构重建。

[ Objective ] To observe the effects of neural stem cells （NSCs） which were differentiated from bone marrow stromal cells （BMSCs） transplantation on the spinal cord injury（SCI） of rats. [ Method] BMSCs were cultured and differentiated to NSCs in vitro. Then the neuron cells were labeled with Brdu （5 -bromodeoxyuridine）. We transplanted the BMSCs into the injured site nine days after the spinal cord injury model was made by Allen device. Sixty adult Wistar rats were randomly divided into three groups ： spinal cord injury cured with transplants of BMSCs to injuryed site （ group A）, spinal cord injury received PBS solution （group B）, control group（group C）. Then the neuron cells labeled with Brdu were detedted by immunohistochemisty, and the reconstruction of spinal cord were detedted by horseradish peroxidase （HRP） staining. [ Result] The neuron cells can be detected in the spinal cord after transplantation. Compared with group B, the number of HRP positive cells of group A is higher. [ Conclusion ] The transplantation of NSCs differentiated from BMSCs can survived in the injuried site and promote the reconstruction. It may be one important approach of repairing the SCI by cells transplantation.