Adherent Cultures of Neural Stem Cells harvested from Subventricular Zone, Subgranular Zone and Spinal cord Ependymal cells in αMEM medium supplemented with 10% FBS

Introduction: After that neural stem cells (NSCs) have been harvested from central nervous system (CNS), rise new hopes in treatment of neurodegenerative disease. NSCs have been harvested from different regions of CNS such as, Subventricular Zone (SVZ) of lateral ventricle, Subgranular Zone (SGZ) of hippocampus and Ependymal cells of spainal cord’s central canal.

The common method for culturing neural stem cells, is suspension culture (neurosphere culture) in specific medium with growth factors. But, neurosphere cultures are often accompanied by progressive loss of self-renewal and differentiation capacity and since the cell populations in neurosphere are heterogeneous, it is hard to determine the quantity and identity of neurosphereforming cells. Therefore, researchers have explored derivation of neural stem cells using adherent cultures.

Expression of Nestin and GFAP is one of neural stem cells traits. Both of these proteins are cytoskeleton microfilaments and expression together just in neural stem cells.

Material and Methods: NSCs have been harvested from SGZ, SVZ and central canal of spinal cord with mechanical and enzymatical digestion and cultured in αMEM medium supplemented with 10% FBS and 1% pen/strip. After 6 days the medium have been changed. In passage 2th and 3th same of the cells have been moved to Petri dishes who are coated with gelatine and examine with immunoreactivity for Nestin and GFAP. Cells have been cultured foe more than three months.

Results: cells harvested from all three regions have been grow in αMEM medium supplemented with 10% FBS and 1% pen/strip and their morphologies was similar to neurons. All three groups were Nestin and GFAP positive.

Conclusion: Results showed NSCs could be cultured in αMEM medium with 10% FBS and in this situation could made a mono layer who adhered to the surface of culture dishes.