中国神经再生研究(英文版) ›› 2021, Vol. 16 ›› Issue (10): 2064-2070.doi: 10.4103/1673-5374.308665

• 原著:神经损伤修复保护与再生 • 上一篇    下一篇

来源于人诱导多能干细胞的神经前体细胞外泌体可改善缺血神经元的功能

  

  • 出版日期:2021-10-15 发布日期:2021-03-19
  • 基金资助:

    浙江省基础公共福利研究计划基金(LGF19H090024);浙江省自然科学基金面上项目(LY17H090006);国家自然科学基金项目(81901073

Exosomes derived from human induced pluripotent stem cell-derived neural progenitor cells protect neuronal function under ischemic conditions

Wen-Yu Li1, #, Qiong-Bin Zhu1, #, Lu-Ya Jin1, Yi Yang1, Xiao-Yan Xu1, Xing-Yue Hu1, 2, *   

  1. 1Department of Neurology, Sir Run Run Shaw Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang Province, China; 2Brain Research Institute, Zhejiang University, Hangzhou, Zhejiang Province, China
  • Online:2021-10-15 Published:2021-03-19
  • Contact: Xing-Yue Hu, MD, PhD, huxingyue2003@zju.edu.cn.
  • Supported by:
    This work was supported by the Foundation of Zhejiang Provincial Basic Public Welfare Research Program of China, No. LGF19H090024 (to XYX); the Natural Science Foundation of Zhejiang Province of China, No. LY17H090006 (to WYL); and National Natural Science foundation of China, No. 81901073 (to QBZ).

摘要:

与其他干细胞相比,来源自人诱导多能干细胞的神经前体细胞在形态和免疫组织学上更类似于皮质神经元,因而在促进神经细胞存活和生长以及缓解星形胶质细胞增生方面具有更大的潜力。有研究表明,移植来源于干细胞外泌体的效果与移植干细胞本身是一致的,均能有效修复神经损伤。但是目前还没有关于源自人诱导多能干细胞的神经前体细胞外泌体对氧糖剥夺神经元的保护的研究。(1)实验首先将胚胎大鼠皮质神经元于无葡萄糖培养基95%N2和5%CO2环境培养1h建立氧糖剥夺模型,然后加入100 µg/mL来源于人诱导多能干细胞的神经前体细胞外泌体培养30min;(2)结果显示,人诱导多能干细胞的神经前体细胞可增加氧糖剥夺神经元的存活率以及培养基中脑源性神经营养因子的水平,抑制氧糖剥夺诱导的神经元中PTEN/AKT信号通路和突触可塑性相关蛋白表达的变化,延长氧糖剥夺神经元最长神经突的长度;(3)验证了来源自人诱导多能干细胞的神经前体细胞的外泌体可通过调节TEN/AKT信号通路和神经突生长,对氧糖剥夺神经元发挥神经保护作用的实验假设。实验于2019年10月10日经邵逸夫医院动物伦理委员会批准(批准号SRRSH20191010)。

https://orcid.org/0000-0002-3590-6801 (Wen-Yu Li); https://orcid.org/0000-0001-6865-6098 (Xing-Yue Hu)

关键词:

多能干细胞, 神经祖细胞, 外泌体, 神经保护, 缺血, PTEN, AKT, 氧糖剥夺, 皮质神经元, 信号通路

Abstract: Compared with other stem cells, human induced pluripotent stem cells-derived neural progenitor cells (iPSC-NPCs) are more similar to cortical neurons in morphology and immunohistochemistry. Thus, they have greater potential for promoting the survival and growth of neurons and alleviating the proliferation of astrocytes. Transplantation of stem cell exosomes and stem cells themselves have both been shown to effectively repair nerve injury. However, there is no study on the protective effects of exosomes derived from iPSC-NPCs on oxygen and glucose deprived neurons. In this study, we established an oxygen-glucose deprivation model in embryonic cortical neurons of the rat by culturing the neurons in an atmosphere of 95% N2 and 5% CO2 for 1 hour and then treated them with iPSC-NPC-derived exosomes for 30 minutes. Our results showed that iPSC-NPC-derived exosomes increased the survival of oxygen- and glucose-deprived neurons and the level of brain-derived neurotrophic factor in the culture medium. Additionally, it attenuated oxygen and glucose deprivation-induced changes in the expression of the PTEN/AKT signaling pathway as well as synaptic plasticity-related proteins in the neurons. Further, it increased the length of the longest neurite in the oxygen- and glucose-deprived neurons. These findings validate the hypothesis that exosomes from iPSC-NPCs exhibit a neuroprotective effect on oxygen- and glucose-deprived neurons by regulating the PTEN/AKT signaling pathway and neurite outgrowth. This study was approved by the Animal Ethics Committee of Sir Run Run Shaw Hospital, School of Medicine, Zhejiang University, China (approval No. SRRSH20191010) on October 10, 2019.

Key words: AKT, cortical neurons, exosome, ischemia, neural progenitor cells, neuronal protection, oxygen and glucose deprivation, pluripotent stem cells, PTEN, signaling pathway

中图分类号: