Passage and Differentiation of Neural Stem Cells from the Hippocampus of the Different Neonate Rats

Objective To investigate the proliferation and differentiation of neural stem cells(NSCs)from hippocampus of neonate rats with different ages.Methods By using trypsin digestion,neural stem cells were isolated from hippocampus of 3-day,10-day and 20-day rats.The medium of serum-free but with basic fibroblast growth factor(bFGF), epidermal growth factor(EGF)and B27 was used to culture NSCs.Immunocytochemistry for nestin was used to identify monoclone NSCs,for glial fibrillary acidic protein(GFAP)and neurone specific enolase(NSE)to identify astrocytes and neurons respectively one week after NSCs were differentiated.The percentage of the NSE positive cells was count and analyzed in statistics.Results The cultured monoclone sphere in 3 groups expressed nestin,and expressed NSE and GFAP one week after induced differentiation.NSCs of 1～6 passages isolated from 3-day rat proliferated much faster than the cells isolated from 10-day and 20-day rats,but almost same after six passage.The number of neurons differentiated from 3d rats was more than that from 10d and 20d rats(P0.05).Conclusion NSCs from 3d rats proliferaterd faster with more number of differentiated neurons compared to 10d and 20d rats.