Effects of primary microglia and astrocytes on neural stem cells in in vitro and in vivo models of ischemic stroke.
Effects of primary microglia and astrocytes on neural stem cells in in vitro and in vivo models of ischemic stroke.
复制标题
原代小胶质细胞和星形胶质细胞对缺血性中风体外和体内模型中神经干细胞的影响
DOI：
10.4103/1673-5374.306093
复制
发表时间：
2021-09
影响因子：
6.1
通讯作者：
Yuan QL
中科院分区：
文献类型：
--
作者：
Wen SJ;Zheng XM;Liu LF;Li NN;Mao HA;Huang L;Yuan QL
神经干细胞(NSCs)移植可以保护动物中风模型中的神经元，但其低存活率和神经元分化限制了其临床应用。神经胶质细胞壁龛是神经干细胞的重要部位，调节神经干细胞的存活、增殖和分化。然而，激活的神经胶质细胞对神经干细胞的影响仍不清楚。在本研究中，我们在体外卒中模型中探讨了激活的星形胶质细胞和小胶质细胞对神经干细胞的影响。我们还研究了神经干细胞和神经胶质细胞联合移植对大鼠卒中后的影响。在Transwell共培养系统中，将原代培养的星形胶质细胞、小胶质细胞或混合胶质细胞暴露在谷氨酸或过氧化氢中，然后种植在上缘，而原代神经干细胞接种在下缘无涂层的孔中，培养7d。结果表明，小胶质细胞有利于神经球的形成，对神经球内的细胞凋亡无影响，而星形胶质细胞和混合胶质细胞则有利于神经球的分化，减少神经球内的细胞凋亡。相反，小胶质细胞和星形胶质细胞在分化培养液中诱导神经干细胞向神经元分化，除了用过氧化氢预处理的星形胶质细胞外，不考虑它们的预处理。采用大脑中动脉结扎法建立大鼠缺血性卒中模型。3d后，将5×105个神经干细胞和小胶质细胞或星形胶质细胞注入右侧脑室。在细胞移植后4天，神经干细胞/星形胶质细胞治疗组大鼠的神经功能缺陷改善情况优于单纯神经干细胞治疗组大鼠。此外，神经干细胞/小胶质细胞和神经干细胞/星形胶质细胞治疗的大鼠与神经干细胞治疗的大鼠相比，缺血体积显著减少。这些结果表明，小胶质细胞和星形胶质细胞对神经干细胞的作用不同，神经干细胞和星形胶质细胞联合移植更有利于缺血性中风大鼠神经功能损害的恢复。该研究于2010年获得同济大学医学院动物伦理委员会中国(批准号：2010-TJAA08220401)的批准。
Transplantation of neural stem cells (NSCs) can protect neurons in animal stroke models; however, their low rates of survival and neuronal differentiation limit their clinical application. Glial niches, an important location of neural stem cells, regulate survival, proliferation and differentiation of neural stem cells. However, the effects of activated glial cells on neural stem cells remain unclear. In the present study, we explored the effects of activated astrocytes and microglia on neural stem cells in vitro stroke models. We also investigated the effects of combined transplantation of neural stem cells and glial cells after stroke in rats. In a Transwell co-culture system, primary cultured astrocytes, microglia or mixed glial cells were exposed to glutamate or H2O2 and then seeded in the upper inserts, while primary neural stem cells were seeded in the lower uncoated wells and cultured for 7 days. Our results showed that microglia were conducive to neurosphere formation and had no effects on apoptosis within neurospheres, while astrocytes and mixed glial cells were conducive to neurosphere differentiation and reduced apoptosis within neurospheres, regardless of their pretreatment. In contrast, microglia and astrocytes induced neuronal differentiation of neural stem cells in differentiation medium, regardless of their pretreatment, with an exception of astrocytes pretreated with H2O2. Rat models of ischemic stroke were established by occlusion of the middle cerebral artery. Three days later, 5 × 105 neural stem cells with microglia or astrocytes were injected into the right lateral ventricle. Neural stem cell/astrocyte-treated rats displayed better improvement of neurological deficits than neural stem cell only-treated rats at 4 days after cell transplantation. Moreover, neural stem cell/microglia-, and neural stem cell/astrocyte-treated rats showed a significant decrease in ischemic volume compared with neural stem cell-treated rats. These findings indicate that microglia and astrocytes exert different effects on neural stem cells, and that co-transplantation of neural stem cells and astrocytes is more conducive to the recovery of neurological impairment in rats with ischemic stroke. The study was approved by the Animal Ethics Committee of Tongji University School of Medicine, China (approval No. 2010-TJAA08220401) in 2010.