Background &Objectives: Stem cells are a class of undifferentiated cells that are able to differentiate into specialized cell types. The discovery of such cells in the adult mammalian central nervous system (CNS), an organ traditionally thought to have little or no regenerative capacity, opened the door to treatment of degenerative diseases of CNS like Stroke, Parkinson, Alzheimer and Spinal Cord Injury. Thus, here we described the isolation of neural stem cells from the adult mouse brain using the neurosphere assay (NSA) and differentiation of these cells to neural adult cells in details.

  Methods: The rostral part of the subventricular zone (SVZ) of the lateral ventricles in the adult mice was dissociated into single cell suspension and cultured using NSA. Primary neurospheres were counted seven days after plating and then the mean number of neurospheres was recorded. The differentiation of neural stem cells into adult neural cells was accomplished by plating the neurosphere-derived cells in differentiating media. Immunocytochemistry and specific markers were used for the identification of the adult neural stem cells.

  Results : The cell suspension obtained from the rostral part of the SVZ of the lateral ventricles generated multipotential colonies, called neurospheres, 7 to 10 days post- incubation. The mean number of neurospheres generated from SVZ was 505±62. The multipotentiality of the neurospheres was shown by palting them in differentiating media and generating adult neural cells including neuron, astrocyte and oligodendrocyte .

  Conclusion: Owing to their rarity and paucity of neural stem cell specific markers, the NSA is a common and selective method for isolating and understanding the biology of embryonic and adult neural stem cells.