大鼠脑出血模型应用骨髓间质干细胞治疗的实验研究

目的 :研究骨髓间质干细胞 (MSCs)移植对脑出血大鼠的治疗效果及机制。方法 :分离MSCs后进行培养、扩增 ,并用流式细胞仪进行荧光三标检测鉴定 ,将标记后的MSCs通过颈动脉、颈静脉、侧脑室 3种途径移植入脑出血大鼠体内 ,用爬行计分法观察大鼠神经功能的改善程度 ,免疫组化法观察在脑内的迁移及分化。结果 :MSCs增殖明显 ,经流式细胞仪检测显示CD90、CD10 6阳性、CD45阴性 ,通过侧脑室、颈动脉移植后大鼠神经功能改善明显 ,移植的MSCs主要迁移到海马区、出血灶等处 ,且分化为神经细胞。结论 :MSCs移植治疗脑出血具有较好的疗效 ,可能分化为神经细胞是其主要机制之一。

The Experimental Research of Therapeutic Efficacy on Mesenchymal Stem Cells(MSCs) Transferring into Model of Rat Brain Following Intracerebral Hemorrhage

Aim:To study therapeutic effect of rat mesenchymal stem cells (MSCs) on intracerebral hemorrhage model and its mechanism in brain.Methods:MSCs were separated from rat bone marrow with percoll (P=1.077 g/L) reagent and culture in culture medium with basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF).MSCs were amplificated until 10 generations and then the fluorescence tri-labelling MSCs were identificated with flow cytometry.Through carotid artery,cervical vein and lateral ventricle MSCs labeling with BrdU were transferred into rat.If the rat neurologic dysfunction ameliorating was observed by behavior scores,MSCs migration and differentiation in brain could be observed by immunohistochemistry and immunofluorescence.Results:MSCs were proliferated with bFGF and EGF and displayed CD90 and CD106 positive but CD45 negative by flow cytometry.The rat neurologic dysfunction was ameliorated significantly by the way of carotid artery and lateral ventricle.The transferred MSCs were mainly migrated to area of hippocampus and hemorrhage.They were also differentiated into nerve cells.Conclusion:The therapy of mesenchymal stem cells transferring into rat brain following intracerebral hemorrhage is effcctive.Probably MSCs differentiating into nerve cells is one of the main mechanisms.