It is now well established that a resident population of neural precursor cells continues to generate new neurons in the adult hippocampus throughout life. Numerous studies have suggested that these newborn neurons preferentially participate in the functional hippocampal circuitry that leads to enhancement of learning, cognition and mood. Therefore, understanding the molecular mechanisms that regulate the activity of these endogenous precursor cells is paramount to develop novel regenerative strategies for the treatment of neurological and psychiatric disorders. The neurosphere assay has been instrumental in discovering the presence of stem and precursor cell population from several brain regions. In this chapter, we describe this assay to specifically isolate and culture neural stem and precursor cell populations from the adult hippocampus of mice. In addition, we provide methods to conduct detailed assays to examine their functional properties such as proliferation, self-renewal, and differentiation.