中国神经再生研究(英文版) ›› 2021, Vol. 16 ›› Issue (4): 653-658.doi: 10.4103/1673-5374.295341

• 原著:神经损伤修复保护与再生 • 上一篇    下一篇

人源多能干细胞可诱导分化为Mu和Kappa阿片受体的神经元

  

  • 出版日期:2021-04-15 发布日期:2020-12-21
  • 基金资助:

    国家自然科学基金(81301065);北京市人才培养计划(D0030340000031)

Neurons derived from human-induced pluripotent stem cells express mu and kappa opioid receptors

Zhi-Hai Ju, Xuan Liang, Yao-Yao Ren, Luo-Wa Shu, Yan-Hong Yan, Xu Cui*   

  1. Department of Anesthesiology, Beijing Tongren Hospital, Capital Medical University, Beijing, China
  • Online:2021-04-15 Published:2020-12-21
  • Contact: Xu Cui, MD, 13141330939@163.com.
  • Supported by:
    This work was supported by the National Natural Science Foundation of China, No. 81301065 (to XC); the Talent Training Plan of Beijing, No. D003034000031 (to XC).

摘要:


脑保护研究显示,诱导多能干细胞的出现为脑保护研究的转化提供了可能。实验使用干细胞培养基培养人源诱导多能干细胞(iPS),设为iPS细胞组;用含有ROCK抑制剂的神经诱导培养基和扩增培养基诱导iPS细胞分化为神经干细胞,进而使用神经元分化培养基和成熟培养基诱导神经干细胞分化为神经元,设为iPS-Neurons组,比较不同分化阶段相关标志物的表达情况。

(1)免疫荧光染色显示:iPS-Neurons组细胞在诱导第11天可表达神经干细胞标志物Sox1、Nestin,诱导第21天可表达神经元标志物TUBB3和NeuN;

(2)PCR结果显示:与iPS细胞组相比,iPS-Neurons组TUBB3基因表达水平上调了15.6倍;

(3)进一步研究发现:与iPS细胞组相比,iPS-Neurons 组神经元Mμ阿片受体(MOR)的基因和免疫反应显著增加(分别为38.3倍和5.7倍),但Kappa阿片受体(KOR)仅略有变化(分别为1.33倍和1.57倍);

(4)上述数据表明:人源iPS细胞能诱导分化为表达MOR和KOR的神经元,可用于体外模拟人类阿片受体功能及潜在机制的研究。


https://orcid.org/0000-0002-2916-790X (Xu Cui) 

关键词: 中风, 脑, 体内, 细胞, 干细胞, 神经分化, 蛋白质, 因子

Abstract: Neuroprotection studies have shown that induced pluripotent stem (iPS) cells have the possibility to transform neuroprotection research. In the present study, iPS cells were generated from human renal epithelial cells and were then differentiated into neurons. Cells in the iPS-cell group were maintained in stem cell medium. In contrast, cells in the iPS-neuron group were first maintained in neural induction medium and expansion medium containing ROCK inhibitors, and then cultivated in neuronal differentiation medium and neuronal maturation medium to induce the neural stem cells to differentiate into neurons. The expression of relevant markers was compared at different stages of differentiation. Immunofluorescence staining revealed that cells in the iPS-neuron group expressed the neural stem cell markers SOX1 and nestin on day 11 of induction, and neuronal markers TUBB3 and NeuN on day 21 of induction. Polymerase chain reaction results demonstrated that, compared with the iPS-cell group, TUBB3 gene expression in the iPS-neuron group was increased 15.6-fold. Further research revealed that, compared with the iPS-cell group, the gene expression and immunoreactivity of mu opioid receptor in the iPS-neuron group were significantly increased (38.3-fold and 5.7-fold, respectively), but those of kappa opioid receptor had only a slight change (1.33-fold and 1.57-fold increases, respectively). Together, these data indicate that human iPS cells can be induced into mu opioid receptor- and  kappa opioid receptor-expressing neurons, and that they may be useful to simulate human opioid receptor function in vitro and explore the underlying mechanisms of human conditions.

Key words: brain, cells, factor, in vivo, neural differentiation, protein, stem cells, stroke