Project Summary Traumatic brain injury (TBI) is a leading cause of death and disability in children with 37,200 children suffering a severe TBI with most dying or experiencing “unfavorable outcomes” within 6 months. Neural stem cells (NSCs) have been seen as a potential therapeutic option for severe pediatric TBI with the ability to differentiate and integrate into TBI brain tissue and replace damaged and lost neural networks. Many of these studies have demonstrated that NSC therapy leads to significant improvements in motor function, learning and memory in rodent models. However, no study to date has closely examined the potential of NSC to undergo region specific differentiation in a discreate injured brain region and correlated it with a measurable change in brain network activity and functional recovery. In this application, we propose to fill this knowledge gap by examining in-depth the effect of transplanted NSC differentiation and engraftment into the damaged primary motor cortex on improving brain motor network activity and motor function in a pediatric piglet model. We hypothesize that induced pluripotent stem cell (iPSC) derived NSCs (iNSCs) transplanted into the damaged primary motor cortex will undergo region specific neural differentiation leading to brain motor network and motor function recovery in a pediatric piglet TBI model. The contribution of this work is highly significant and innovative since successful completion of this project will directly lead to new insights into the ability of transplanted iNSC to undergo regionalized specific differentiation and integration, and rigorously and quantitatively assess functional recovery at the brain and whole animal level. This critical information will advance TBI cell therapy with broad implications in the neural injury, cell therapy, and tissue engineering fields at large. Testing in a translational pig model is critical due to key similarities in brain anatomy and physiology relative to rodents; thus, making findings in the pig more predictive of what would occur in human patients.