脑皮层创伤灶注射法移植神经干细胞的实验研究

Empirical study on transplantation of neural stem cells into traumatic lesions in the cerebral cortex by injection method

目的观察食蟹猴自体骨髓源神经干细胞（NSCs）移植到皮层创伤灶内的存活、生长情况。方法取骨髓分离、纯化骨髓基质细胞，培养诱导成NSCs，Nestin染色阳性说明具备NSCs特征。再加入细胞标记物BrdU进行共培养7d待移植之用。移植前将BrdU标记的NSCs收集、离心，制成干细胞悬液。食蟹猴分即时移植和延迟移植组，用微量注射针将NSCs悬液注入到猴脑皮质损伤灶及创面周边脑皮层，移植后1，3，6个月各灌杀2只食蟹猴，做移植区组织切片染色检查。结果即时移植组和延迟移植组都可观察到脑皮层创伤灶内有BrdU标记阳性细胞，而假移植组织切片中则未见BrdU阳性细胞。移植后1，3，6个月染色可见移植细胞早期呈簇状分布，移植后6个月在移植区邻近的脑白质内也可观察到有BrdU阳性细胞，正电子发射电子计算机断层扫描（PET）检查显示移植NSCs后创伤区域脑组织葡萄糖代谢有所恢复。结论移植的骨髓源性NSCs在脑内有存活，并向邻近区域迁移，有助于创伤脑组织修复。

Objective To observe the survival and growth of autologous bone marrow-derived neural stem cells （BMdNSCs） after transplantation into cerebral cortex traumatic lesions in long-tailed macaques. Methods Long-tailed macaque BMdNSCs characterized by Nestin staining were prepared and cultured with cell marker BrdU for 7 days. BMdNSCs were then collected and suspended. BMdNSC suspension was injected immediately （immediate transplantation group, Group A） and later （delayed transplantation group, Group B） into the traumatic lesions in the cerebral cortex with micro-injection needle. Two animals were sacrificed at 1, 3 or 6 months and tissue sections from the transplantation area were stained. Results In Groups A and B, positive BrdU cells were observed in the lesions, while no positive BrdU cells were found in sham transplantation group. At 1,3 and 6 months after transplantation, transplanted BMdNSCs were distributed at solitary positions. At 6 months, positive BrdU cells were also observed in adjacent white matter, and PET examination showed a recovery of glucose metabolism in the lesions. Conclusion Bone marrow-derived neural stem cells can survive in the brain and migrate to the injured brain areas, which may contribute to tissue repair.