Isolation, Cultivation, Differentiation And Identification Of Neural Stem Cells From Neonatal SD Rat Hippocampus

The conventional idea is that the adult mammal central nervous system could not regeneration , which means the neurons of the adult brain and spinal cord will decrease because of trauma, disease and apoptosis et al. The discovery of neural stem cells throws off the traditional idea. The studies of last decade revealed that there are neural stem cells in some places of adult central nervous system. In serum-free medium, stimulated by nutrition factors, the neural stem cells can proliferate and form neurospheres. They can serial passage and differentiate into neurons , astrocytes and oligodendrocytes . In vivo , the stem cells are in resting in normal condition. They can proliferate and differentiate into neurons or glial cells post trauma. The animal experiments of neural stem cells transplantation reveal: The transplanted neural stem cells can live, immigrate and differentiate into glial cells and neurons. Some neurons form functionally synapse with the host cells, and the functions are improved significantly. The researchment of neural stem cells had already become into the front edge in the field of neuroscience.Objective : Isolation of neural stem cells from the neonatal rat hippocampus for further cultivation, differentiation and identification.Methods: Hippocampi of neonatal rats (within 24 hours)were taken out and mechanically dissociated into single cell suspension. After serum-free cultivation and fetal bovine serum-induced proliferation, neural stem cells and their progeny cells were identified by immunofluorescence technique.Result: The cells isolated from the hippocampus of neonatal rat were nestin immune-positive, and could be continuously passage cultured in vitroand form clone. The cells after differentiation could express the specific antigens NeuN and glial fibrillary acidic protein in neuron and astrocyte respectively.conclusions: The isolated and cultured cells can express specific marker nestin of neural stem cells ,and have strong capability of proliferation and pluripotent differentiation. It is suggested that neural stem cells exist in neonatal rat hippocampus and can be used for further experimental studies.