In hypoxic-ischemic encephalopathy, the neural progenitors (NPs) of the developing brain fail to replenish the oligodendrocyte progenitor cells lost during hypoxic-ischemic injury (HII). In this study, we aim to examine the influence of HII on the vulnerability of human NPs derived from human embryonic stem cells with regard to cell survival and oxidative stress, followed by assessment of cellular deregulation through measuring glutathione levels, basal calcium, glutamate release, and intracellular calcium ([Ca²⁺]_i) response under KCl and ATP stimulation. NPs were further evaluated for their fundamental potential of self-renewal and proliferation, neural and glial progenitor pool, and migration. Oxygen-glucose deprivation (OGD) of 90 min was sublethal for NPs, yet significantly increased reactive oxygen species generation and oxidative stress susceptibility, and decreased glutathione levels, along with a rise in glutamate release, basal [Ca²⁺]_i, and KCl and ATP-induced [Ca²⁺]_i. Distinct increase in gene expression for K⁺ leak channel (Twik-related acid-sensitive K⁺ channel 1 [TASK-1]) and purinergic receptor P2X7, and decrease of voltage-gated K_v channels K_v1.5, K_v4.2, and K_v4.3 were observed. TASK-1 increase was detected by FACS too. OGD-insulted NPs showed reduced migration potential and decline in glial progenitor population. This study thus demonstrates for the first time that brief exposure of OGD does not reduce the NP population, its proliferation, and self-renewal, but can induce significant alteration in oxidative stress susceptibility, glutamate release, [Ca²⁺]_i response to physiological stimulus, migration, and glial progenitor pool. We thus infer that treatment strategies need to target repair of NPs of the developing brain that is affected during intrapartum asphyxia, leading to varying neurologic complications such as seizure, mental retardation, and/or cerebral palsy.

中文翻译：

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使用源自人类胚胎干细胞的神经祖细胞评估体外新生儿缺氧缺血性损伤。

在缺氧缺血性脑病中，发育中大脑的神经祖细胞 (NPs) 无法补充缺氧缺血性损伤 (HII) 期间丢失的少突胶质祖细胞。在这项研究中，我们旨在检查 HII 对源自人类胚胎干细胞的人类 NPs 在细胞存活和氧化应激方面的脆弱性的影响，然后通过测量谷胱甘肽水平、基础钙、谷氨酸释放、和细胞内钙（[Ca ²⁺ ] _i) 在 KCl 和 ATP 刺激下的反应。NPs 的自我更新和增殖、神经和神经胶质祖细胞池以及迁移的基本潜力被进一步评估。90 分钟的氧-葡萄糖剥夺 (OGD) 对 NPs 是亚致死的，但显着增加了活性氧的产生和氧化应激敏感性，并降低了谷胱甘肽水平，同时谷氨酸释放、基础 [Ca ²⁺ ] _i和 KCl 增加和 ATP 诱导的 [Ca ²⁺ ] _i。K ⁺泄漏通道（Twik 相关酸敏感 K ⁺通道 1 [ TASK-1 ]）和嘌呤能受体P2X7 的基因表达明显增加，并观察到电压门控 K _v通道K _v 1.5、K _v 4.2和K _v 4.3的减少。FACS 也检测到 TASK-1 增加。OGD 损伤的 NPs 显示迁移潜力降低和神经胶质祖细胞数量下降。因此，本研究首次证明，短暂暴露于 OGD 不会减少 NP 种群、其增殖和自我更新，但可以诱导氧化应激易感性、谷氨酸释放、[Ca ²⁺ ] _{i 的}显着改变。对生理刺激、迁移和胶质祖细胞池的反应。因此，我们推断治疗策略需要针对在产时窒息期间受影响的发育中大脑的 NPs 的修复，导致各种神经系统并发症，如癫痫发作、智力低下和/或脑瘫。