定向诱导的骨髓基质细胞移植后在大鼠缺血脑组织中的分布

Study of transplantion of human bone marrow stromal cells after orientiatedly induced into neuron-like cells

目的探讨人骨髓基质细胞(bone marrow stromal cells,MSCs)源神经细胞体内移植后的分布状况。方法采用密度梯度离心结合贴壁分离法分离、纯化人MSCs。MSCs经bFGF预诱导24h及阿魏酸钠(Sodium Ferulate)诱导24h后用于移植。采用线栓法复制大鼠局灶性脑缺血再灌注动物模型,缺血2h后行再灌注,24h后经栓塞侧颈内动脉注入诱导后的MSCs(5×106/0.8ml)(n=6)。2w后取脑组织行冰冻切片,通过检测人细胞核特异抗体MAB1281来鉴定移植的人MSCs。结果MSCs经阿魏酸钠诱导后,细胞逐渐圆缩并伸出突触,呈现神经细胞样形态。在移植大鼠脑内可检测到MAB1281呈阳性表达的人MSCs,且MSCs主要分布在缺血侧损伤灶的周围,明显高于对侧同部位及其它脑区。结论人MSCs源神经细胞移植后在大鼠脑内存活并主要聚集在脑内缺血灶的周围。

Objective To investigate the distribution of human bone marrow stromal cells（MSCs） transplanted into rat brain after induced into neuron-like cells. Methods MSCs were isolated by gradient centrifugation in lymphocyte separation medium and by their adherence to tissue culture surfaces. The cultured MSCs were induced with sodium ferulate （SF） for 24 hours after pre-induced by bFGF for 24 hours. The right middle cerebral artery occlusion （MCAO） model of SD rats was produced by obstructing MCA using a single strand nylon thread, and we reperfused the artery after 2 hours. The induced MSCs about 5×10^6/0.8ml were transplanted into the right internal carotid artery of MCAO rats after reperfusion for 24 hours （n=6）. After 2 weeks, frozen sections of rat brain were made paralleling the coronal plane. The expression of MAB1281 in the brain tissue was detected by immunofluorescence staining. Results 24 hours after the MSCs induced by SF, cell morphological changes occurred, which extended long processes as the cellular body contracted. The positive cells of MAB1281 were observed in the same regions of brain surround the ischemic focus by immunofluorescence staining. Conclusion MSCs induced by SF can survive and aggregated around the ischemic zone in the brain of MCAO rats.