中国神经再生研究(英文版) ›› 2015, Vol. 10 ›› Issue (9): 1516-1522.doi: 10.4103/1673-5374.165526

• 原著:神经损伤修复保护与再生 • 上一篇    下一篇

5%O2预处理72 h为促进大脑皮质神经干细胞增殖及分化的最佳浓度和时间窗

  

  • 收稿日期:2015-07-12 出版日期:2015-09-28 发布日期:2015-09-28
  • 基金资助:

    济宁市科技局项目(2012jnjc07)

Optimal concentration and time window for proliferation and differentiation of neural stem cells from embryonic cerebral cortex: 5% oxygen preconditioning for 72 hours

Li-li Yuan1, 2, Ying-jun Guan2, Deng-dian Ma3, Hong-mei Du2   

  1. 1 Department of Histology and Embryology, Academy of Basic Medicine, Jining Medical University, Jining, Shandong Province, China
    2 Department of Histology and Embryology, Academy of Basic Medicine, Weifang Medical University, Weifang, Shandong Province, China
    3 Department of Otorhinolaryngology, Affiliated Hospital of Jining Medical University, Jining, Shandong Province, China
  • Received:2015-07-12 Online:2015-09-28 Published:2015-09-28
  • Contact: Li-li Yuan or Ying-jun Guan, Ph.D., liliyuan06@126.com or guanyj@wfmc.edu.cn.
  • Supported by:

    This work was supported by the Science Foundation of Jining Science and Technology Bureau of China, No. 2012jnjc07.

摘要:

低氧可以促进胚胎12 d大鼠脑组织神经干细胞增殖及分化,但寻找恰当的有利于神经干细胞增殖及分化的低氧预处理的浓度和时间目前尚存在争议。为了解决此问题,我们体外培养了SD大鼠14 d胚胎脑皮质神经干细胞,分别在5%和10%O2低氧预处理条件下进行培养观察。MTT实验、神经球计数检测、免疫荧光染色结果显示,5%或10%O2低氧预处理72 h均能提高大鼠胚胎脑皮质神经干细胞的活性及增殖能力,但当低氧时间达到120 h,神经干细胞凋亡比例增加;5%O2低氧预处理72 h在促进脑皮质神经干细胞增殖的同时,还能同时促进其向神经元分化。结果证实,低氧预处理促进大脑皮质神经干细胞的增殖及分化的最佳干预浓度为5%O2,最佳干预时间窗为72 h。

关键词: 神经再生, 脑损伤, 神经干细胞, 低氧, 脑皮质, 凋亡, 分化, 大鼠, 微管相关蛋白2, 胶质纤维酸性蛋白, caspase-3

Abstract:

Hypoxia promotes proliferation and differentiation of neural stem cells from embryonic day 12 rat brain tissue, but the concentration and time of hypoxic preconditioning are controversial. To address this, we cultured neural stem cells isolated from embryonic day 14 rat cerebral cortex in 5% and 10% oxygen in vitro. MTT assay, neurosphere number, and immunofluorescent staining found that 5% or 10% oxygen preconditioning for 72 hours improved neural stem cell viability and proliferation. With prolonged hypoxic duration (120 hours), the proportion of apoptotic cells increased. Thus, 5% oxygen preconditioning for 72 hours promotes neural stem cell proliferation and neuronal differentiation. Our findings indicate that the optimal concentration and duration of hypoxic preconditioning for promoting proliferation and differentiation of neural stem cells from the cerebral cortex are 5% oxygen for 72 hours.

Key words: nerve regeneration, brain injury, neural stem cells, low oxygen, cerebral cortex, apoptosis, differentiation, microtubule-associated protein 2, glial fibrillary acidic protein, caspase-3, neural regeneration