Ju Zhi-Hai, Liang Xuan, Ren Yao-Yao, Shu Luo-Wa, Yan Yan-Hong, Cui Xu
Department of Anesthesiology, Beijing Tongren Hospital, Capital Medical University, Beijing, China.
Neural Regen Res. 2021 Apr;16(4):653-658. doi: 10.4103/1673-5374.295341.
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.
神经保护研究表明,诱导多能干细胞(iPS细胞)有可能改变神经保护研究。在本研究中,iPS细胞由人肾上皮细胞生成,然后分化为神经元。iPS细胞组的细胞维持在干细胞培养基中。相比之下,iPS神经元组的细胞首先维持在含有ROCK抑制剂的神经诱导培养基和扩增培养基中,然后在神经元分化培养基和神经元成熟培养基中培养,以诱导神经干细胞分化为神经元。在分化的不同阶段比较相关标志物的表达。免疫荧光染色显示,iPS神经元组的细胞在诱导第11天表达神经干细胞标志物SOX1和巢蛋白,在诱导第21天表达神经元标志物TUBB3和NeuN。聚合酶链反应结果表明,与iPS细胞组相比,iPS神经元组中TUBB3基因表达增加了15.6倍。进一步研究表明,与iPS细胞组相比,iPS神经元组中μ阿片受体的基因表达和免疫反应性显著增加(分别为38.3倍和5.7倍),但κ阿片受体的基因表达和免疫反应性仅有轻微变化(分别增加1.33倍和1.57倍)。总之,这些数据表明人iPS细胞可被诱导为表达μ阿片受体和κ阿片受体的神经元,它们可能有助于在体外模拟人阿片受体功能并探索人类疾病的潜在机制。