Whereas neural stem cells and their niches have been extensively studied in the brain, little is known on these cells, their environment and their function in the adult spinal cord. Adult spinal cord neural stem cells are located in a complex niche surrounding the central canal and these cells expressed genes which are specifically expressed in the caudal central nervous system (CNS). In depth characterization of these cells in vivo and in vitro will provide interesting clues on the possibility to utilize this endogenous cell pool to treat spinal cord damages. We describe here a procedure to derive and culture neural spinal cord stem cells from adult mice using the neurosphere method.