Ischemic stroke is one of the most serious and widespread neurological emergencies in the world. It is characterized by rapid disruption of brain blood flow, and it can cause irreversible damage to brain cells leading to permanent disability or death. Currently, treatment options are limited and fail to completely prevent or reverse brain damage. It is therefore essential to pursue innovative approaches to improve the prognosis and quality of life of ischemic stroke patients.Brain organoids are three-dimensional models that can reproduce the cellular composition, structure, and neuronal connectivity of the human brain.These organoids thus offer a promising opportunity to develop a model of ischemic stroke. In this work, we explore the use of hiPSC brain organoids as a model for studying ischemic stroke.Organoids were produced at different times to define which is the best time point to mimic the ischemic human brain. For this purpose, we performed western blotting, imaging, dPCR, calcium activity assay, and electrophysiological evaluation with microelectrode array. The ischemic stroke was modeled with oxygen-glucose deprivation (OGD) at different times (2h – 8h). Viability and propidium iodide assays were performed to evaluate the efficacy of the OGD.Organoids at 50days of culture were chosen as they present a complete neuronal maturation and presence of the glia. The vitality experiment showed that the longer the OGD the more the organoids were damaged.In conclusion, this study provides a humanized ischemic stroke model represented by brain organoids that will be used as a new platform for drug testing.