Establishing a stoke experimental model, which is better in line with the physiology and function of human brain, is the bottleneck for the development of effective anti-stroke drugs. A three-dimensional cerebral organoids (COs) from human pluripotent stem cells can mimic cell composition, cortical structure, brain neural connectivity and epigenetic genomics of in-vivo human brain, which provides a promising application in establishing humanized ischemic stroke model. COs have been used for modeling low oxygen condition-induced hypoxic injury, but there is no report on the changes of COs in response to in vitro oxygen-glucose deprivation (OGD)-induced damage of ischemic stroke as well as its application in testing anti-stroke drugs. In this study we compared the cell composition of COs at different culture time and explored the cell types, cell ratios and volume size of COs at 85 days (85 d-CO). The 85 d-CO with diameter more than 2 mm was chosen for establishing humanized ischemic stroke model of OGD. By determining the time-injury relationship of the model, we observed aggravated ischemic injury of COs with OGD exposure time, obtaining first-hand evidence for the damage degree of COs under different OGD condition. The sensitivity of the model to ischemic injury and related treatment was validated by the proven pan-Caspase inhibitor Z-VAD-FMK (20 μM) and Bcl-2 inhibitor navitoclax (0.5 μM). Neuroprotective agents edaravone, butylphthalide, P7C3-A20 and ZL006 (10 μM for each) exerted similar beneficial effects in this model. Taken together, this study establishes a humanized ischemic stroke model based on COs, and provides evidence as a new research platform for anti-stroke drug development.

建立更符合人脑生理和功能的stoke实验模型是开发有效抗脑卒中药物的瓶颈。来自人类多能干细胞的三维大脑类器官 (COs) 可以模拟体内人脑的细胞组成、皮质结构、脑神经连接和表观遗传基因组学，这为建立人源化缺血性中风模型提供了有希望的应用。COs已被用于模拟低氧条件下引起的缺氧损伤，但尚无关于COs响应体外氧-葡萄糖剥夺（OGD）诱导的缺血性脑卒中损伤的变化及其在测试抗损伤中的应用的报道。 - 中风药物。在这项研究中，我们比较了 CO 在不同培养时间的细胞组成，并探索了细胞类型，85 天 (85 d-CO) 时 CO 的细胞比率和体积大小。选择直径大于2mm的85d-CO建立OGD缺血性卒中人源化模型。通过确定模型的时间-损伤关系，观察COs随OGD暴露时间的加重缺血性损伤，获得不同OGD条件下COs损伤程度的第一手证据。该模型对缺血性损伤和相关治疗的敏感性已通过成熟的泛半胱天冬酶抑制剂 Z-VAD-FMK (20 μM) 和 Bcl-2 抑制剂 navitoclax (0.5 μM) 进行验证。神经保护剂依达拉奉、丁苯酞、P7C3-A20 和 ZL006（各 10 μM）在此模型中发挥了类似的有益作用。综上所述，本研究建立了基于 COs 的人源化缺血性卒中模型，