摘要
背景:脑缺血导致神经功能障碍的根本原因是神经元损伤,促进神经元再生是神经功能恢复的关键,而内源性神经干细胞的修复作用有限,外源性神经干细胞移植为脑缺血后组织重构与功能恢复带来希望,但种子细胞的选择仍缺乏系统的比较研究。目的:比较3种不同来源神经干细胞移植治疗大鼠脑缺血损伤的效果。方法:分离培养SD大鼠脂肪间充质干细胞、骨髓间充质干细胞和皮肤来源的诱导性多能干细胞,在体外进行神经向诱导分化7 d检测神经干细胞的转化率。取脑缺血损伤模型造模成功的100只SD大鼠,随机分为模型组、骨髓间充质干细胞来源组、诱导性多能干细胞来源组、脂肪间充质干细胞来源组和磷酸盐缓冲液组,每组20只,移植相应的细胞悬液或磷酸盐缓冲液;另随机取20只SD大鼠为假手术组,20只SD大鼠为对照组,不进行细胞移植。细胞移植后1周和4周进行各项指标检测。结果与结论:(1)体外诱导分化7 d后,诱导性多能干细胞来源的神经干细胞的转化率明显高于脂肪间充质干细胞和骨髓间充质干细胞,差异有显著性意义(P<0.01);(2)移植1周和4周后,移植不同来源神经干细胞的大鼠神经元凋亡数量和脑梗死体积明显小于磷酸盐缓冲液组和模型组(P<0.05),诱导性多能干细胞来源组神经功能缺损评分、脑梗死体积和神经元凋亡数量明显低于脂肪间充质干细胞来源组和骨髓间充质干细胞来源组(P<0.05);(3)移植4周时,各组大鼠神经功能缺损评分、神经元凋亡数量和脑梗死体积明显低于移植1周(P<0.01);(4)移植神经干细胞3组大鼠纹状体神经元胞质内尼氏体数量较对照组和假手术组明显增多;(5)研究结果表明,3种来源的神经干细胞均能明显改善神经缺损症状、缩小缺血区域的脑梗死面积、减少神经元变性和凋亡的数量,特别是诱导性多能干细胞具有更高的分化能力和更好的移植效果。
BACKGROUND： The underlying mechanism of neurological dysfunction caused by cerebral ischemia is neuronal damage, and facilitating neuronal regeneration is the key to neural function recovery. However, endogenous neural stem cells have limited use in nerve repair. Transplantation of exogenous neural stem cells brings hope for tissue remodeling and functional recovery after cerebral ischemia, but systematic comparative studies on the selection of seed cells are still lacking.OBJECTIVE： To compare the therapeutic effect of neural stem cells from three sources in the treatment of cerebral ischemia injury in rats.METHODS： Adipose-derived mesenchymal stem cells （ADMSCs）, bone marrow mesenchymal stem cells （BMSCs）, and skin-derived induced pluripotent stem cells （iPSCs） from Sprague-Dawley rats were isolated and cultured, and induced in vitro to differentiate into neural stem cells. Conversion efficiency was detected at 7 days of neuronal induction. One hundred rat models of cerebral ischemia were randomized into model, BMSCs, iPSCs, ADMSCs, and PBS groups, with 20 rats in each group. Then, corresponding cell suspension or PBS solution was transplanted in each group. Another 40 Sprague-Dawley rats were taken as sham operation group （n=20） and control group （n=20） with no intervention. Index measurements were performed at 1 and 4 weeks after cell transplantation.RESULTS AND CONCLUSION： After 7 days of in vitro induction, the conversion efficiency of iPSCs was significantly higher than that of ADMSCs and BMSCs （P 〈 0.01）. At 1 and 4 weeks after transplantation, neuronal apoptosis and infarction size were significantly reduced in the iPSCs, ADMSCs and BMSCs groups as compared with the PBS and model groups （P 〈 0.05）. Moreover, neurological severity score, infarction size and neuronal apoptosis were significantly reduced in the iPSCs group as compared with the ADMSCs and BMSCs groups （P 〈 0.05）. At 4 weeks after cell transplantation, neurological severity score, infarction size and neuronal apoptosis in each group showed significant reduction as compared with the values at 1 week after cell transplantation （P 〈 0.01）. Numbers of Nissl bodies in neurons of the striatum in the three cell transplantation groups were significantly higher than those in the control and sham operation groups. Our findings from this study reveal that neural stem cells differentiated from three sources can significantly improve the symptoms of neurological deficits, and reduce infarction size, neuronal degeneration and neuronal apoptosis. iPSCs especially have better differentiation ability and therapeutic effects than BMSCs and ADMSCs.
作者
钟波
柳霞
吴伟
Zhong Bo;Liu Xia;Wu Wei(Department of Neurology;Department of Electrocardiogram,Linqing People＇s Hospital,Linqing 252600,Shandong Province,China;Department of Neurology,Qilu Hospital of Shandong University,Jinan 250012,Shandong Province,China)
出处
《中国组织工程研究》
CAS
北大核心
2018年第21期3349-3356,共8页
Chinese Journal of Tissue Engineering Research
