Our research is focused on studying how the neural stem cell population is regulated in the adult mammalian brain. Understanding the intrinsic and extrinsic molecular mechanisms that regulate neural stem cells within the neurogenic niches is important to comprehend the biology and to develop future potential therapeutic applications. To study neural stem cell behavior, both in vivo and in vitro approaches are widely used.
In vitro neural stem cell cultures offer controlled environments and the possibility to easily manipulate and monitor this population of cells. Techniques for gene expression preservation in vitro are a versatile approach to study molecular mechanisms governing cell biology. However, an efficient and reproducible method for overexpressing and knocking down candidate genes in neural stem cells is still challenging in the field.
Traditional transfection methods for gene delivery have proven effective in central nervous system cells. Moreover, these methods affect cell viability and functionality. Thus, refinement of alternative approaches is crucial to manipulate gene expression in neural stem cells.
With this protocol, we present an improved nucleofection system that achieves high efficiency of gene delivery in neural stem cell cultures from the adult murine subventricular zone, along with survival rates higher than 80%
