Stroke is a devastating neurological disorder and one of the leading causes of mortality and disability. To understand the cellular and molecular mechanisms of stroke and to develop novel therapeutic approaches, two different in vitro human cell-based stroke models were established using oxygen-glucose deprivation (OGD) conditions. In addition, the effect of adipose stem cells (ASCs) on OGD-induced injury was studied. In the present study, SH-SY5Y human neuroblastoma cells and human induced pluripotent stem cells (hiPSCs) were differentiated into neurons, cultured under OGD conditions (1% O₂) for 24 h, and subjected to a reperfusion period for 24 or 72 h. After OGD, ASCs were cocultured with neurons on inserts for 24 or 72 h to study the neuroprotective potential of ASCs. The effect of OGD and ASC coculture on the viability, apoptosis, and proliferation of and axonal damage to neuronal cells was studied. The results showed that OGD conditions induced cytotoxicity and apoptosis of SH-SY5Y- and hiPSC-derived neurons, although more severe damage was detected in SH-SY5Y-derived neurons than in hiPSC-derived neurons. Coculture with ASCs was protective for neurons, as the number of dead ASC-cocultured neurons was lower than that of control cells, and coculture increased the proliferation of both cell types. To conclude, we developed in vitro human cell-based stroke models in SH-SY5Y- and hiPSC-derived neurons. This was the first time hiPSCs were used to model stroke in vitro. Since OGD had different effects on the studied cell types, this study highlights the importance of using several cell types in in vitro studies to confirm the outcomes of the study. Here, ASCs exerted a neuroprotective effect by increasing the proliferation and decreasing the death of SH-SY5Y- and hiPSC-derived neurons after OGD.

中文翻译：

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人神经母细胞瘤细胞和诱导多能干细胞衍生神经元的体外氧-葡萄糖剥夺诱导的中风模型。

中风是一种毁灭性的神经系统疾病，是导致死亡和残疾的主要原因之一。为了了解中风的细胞和分子机制并开发新的治疗方法，使用氧-葡萄糖剥夺（OGD）条件建立了两种不同的体外人细胞为基础的中风模型。此外，研究了脂肪干细胞（ASCs）对OGD诱导的损伤的作用。在本研究中，SH-SY5Y人神经母细胞瘤细胞和人诱导性多能干细胞（hiPSC）分化为神经元，并在OGD（1％O ₂）24小时，然后进行24或72小时的再灌注。OGD后，将ASC与插入物上的神经元共培养24或72 h，以研究ASC的神经保护潜力。研究了OGD和ASC共培养对神经元细胞活力，凋亡和轴突损伤的影响。结果显示，尽管SH-SY5Y和hiPSC衍生的神经元比hiPSC衍生的神经元检测到更严重的损伤，但是OGD条件诱导了SH-SY5Y和hiPSC衍生的神经元的细胞毒性和凋亡。与ASC的共培养对神经元具有保护作用，因为死亡的ASC共培养的神经元数量少于对照细胞，并且共培养增加了两种细胞类型的增殖。总而言之，我们在体外进行了开发SH-SY5Y和hiPSC衍生的神经元中基于人细胞的中风模型。这是首次使用hiPSC在体外模拟中风。由于OGD对所研究的细胞类型具有不同的影响，因此本研究强调了在体外研究中使用几种细胞类型来确认研究结果的重要性。在这里，ASC通过增加OGD后SH-SY5Y和hiPSC衍生的神经元的增殖并减少其死亡而发挥神经保护作用。