Background: Ischemic stroke increases the number of glial cells, such as astrocytes, and causes neuronal death, disrupting the neuron-to-glia balance, contributing to neurodegeneration. Treatment with NeuroD-adeno-associated virus (NeuroD1-AVV) may enhance neuronal transdifferentiation and improve motor function, but the optimal administration protocol for the drug has yet to be determined. Methods: Non-human primates (NHPs) underwent middle cerebral occlusion surgery. Fourteen days poststroke, subjects received NeuroD1-AVV according to two distinct protocols: Three high doses and three low doses. Neurological deficits and cognitive performance were measured using the NHP stroke scale and coloured glove shift of set task, respectively. Nine months post-stroke, NHPs were euthanized. Brains were harvested and stained for neuronal (NEUN and MAP2) and glial (GFAP, IBA1) markers using immunofluorescence techniques. Results: Our results indicate that both protocols effectively rebalance the neuron-to-glia cell ratio by decreasing GFAP+ cells in the P1 and P2 NHPS ipsilateral hemispheres. No cognitive performance differences were found across groups; however, P2 had better NHPSS outcomes from months 2 to 9. Conclusions: The findings support both injection protocols in restoring histological balance, with P2 being more effective for motor function rehabilitation. Investigations into neuronal functionality and development levels continue.