猴脑血管缺血模型复制及骨髓间充质干细胞移植治疗

Establishment of Ischemic Cerebral Infarction Model and the Treatment with Bone Marrow Mesenchymal Stem Cells in Monkey

目的建立猕猴局灶性血管缺血性脑损伤模型,评价自体骨髓间充质干细胞移植治疗脑损伤的疗效及临床治疗可行性。方法用导管介入法从股动脉将栓子输送至大脑中动脉使大脑中动脉分支动脉栓塞,诱发局灶性缺血性脑损伤,经CT扫描确认脑损伤及部位和行为学观察符合脑损伤要求。分离扩增自体骨髓间充质干细胞至第4代,于脑血管栓塞后30d,在CT引导下将4×108细胞/kg骨髓间充质干细胞移植到脑损伤部位,每月1次连续观察脑损伤面积及行为学变化。结果脑血管栓塞前造影明显可见左侧大脑中动脉血管树及其分支分布情况,栓塞后血管造影证实额颞区无血管显示,随后连续CT扫描见额颞叶区明显梗死灶,行为学观察可见血管栓塞对侧前后肢运动功能严重障碍。间充质干细胞移植后3个月可见脑损伤面积明显缩小,脑神经功能明显改善,半年后脑损伤面积进一步缩小,脑神经功能基本恢复正常。结论自体骨髓间充质干细胞移植有助于缺血性脑损伤结构修复和功能恢复。

Objective To establish a monkey ischemic cerebral infarction model and to confirm the therapeutic value of mesenchymal stem cells transplantation for treatment of cerebral infarction. Methods 10 male monkeys of 8-10 years old were used in this study.The branches of middle cerebral artery were thrombed by injecting with PVA. The blood vessel blockage and brain damage were evaluated by middle cerebral artery digital subtraction angiography and CT scanning before and after cerebral infarctions. Marrow mesenchymal stem cells（MMSCs） were induced to express nestin by NGF and 4×10^8 cell/kg induced cells were injected into the boarder area of the damaged brain tissue after one month of vascular embolization. Standard assessment of neural function was performed to evaluate the neural function deficit. The pathological change of infracted brain tissue was tested by CT scanning monthly. Results Thrombosis of middle cerebral artery branch was confirmed by DSA after PVA injection. Cerebral infarction was found in the area of the middle cerebral artery from 12 hours to one month after vascular embolization. The appearance of brain lesion coincides with the defect （grade 3） of neural function. Significant change of brain lesion and neural function were not found in the control monkey, while brain tissue damage area reduced greatly and neural functioh improved in the monkey after transplantation. Conclusion A monkey model of ischemic cerebral infarction was established by injecting PVA using the methods of artery interventional procedures. The appearance of brain lesion and neural function can be reversed greatly by transplanting the nestin positive cells derived from MMSCs. The results indicated that nestin positive ceils from MMSCs can be used to treat stroke, ischemic cerebral infarction and other brain damaged diseases.