روش جديد و كار آمد براي توليد نوروسفر بيشتر از بطن طرفي مغز موش بالغ

A New and Effective Method for More Neurosphere Forming from the Adult Mouse Brain Lateral Ventricle

Background & Objective: It is now clear that the adult mammalian subventricular zone (SVZ) contains a population of neural stem cells (NSCs) that give rise to neurons and glia. Owing to their rarity, and a paucity of NSC-specific markers, the neurospher assay (NSA) is a common and selective method for isolating and understanding the biology of embryonic and adult neural stem cells. There are different methods for neurosphere growing from different regions of the CNS including Lateral ventricles. The objective of this study is introducing a new and effective strategy for more neurosphere firming from the SVZ of the adult mouse brain lateral ventricle using NSA. Methods: Two different methods were used in order to isolate and culture the SVZ of the lateral ventricles using NSA. In the first method (Ritze and Reynolds method) the rostral part of the SVZ of the lateral ventricles was dissected into single cell suspension and cultured using NSA. In the second method (vibratome resecting of the brain) after cutting the brain into 400 pim serial sections using a vibratom, the SVZ was microdissected from all sections of rostral part of lateral ventricle and cultured separately, using NSA. Primary neurospheres were counted seven days after plating. Then the mean numbers of neurospheres generated in two different methods were compared. Results: The mean number of neurospheres generated by sectioning method was much higher than the one generated using first method (P<0.0001). The distribution and frequency of neurosphere forming cells (or NSCs) is not the same along the antero-posterior axis of the rostral part of the lateral ventricle. The greatest frequency of neurosphere forming cells was detected in 0.74mm rostral to the bregma. Conclusion: Second section method, due to more neurosphere generation, in comparison with the first method is more appropriate and efficient for neurosphere forming from the SVZ of the lateral ventricle.