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[Isolation, culture and identification of rat hippocampal neural stem cells]
1Department of High-temperature Medicine, Southern Medical University, Guangzhou 510515, China.
Objective:
To explore the feasibility of the isolation, culture and identification of the neural stem cells originated from the hippocampi of neonatal rats (1-3 d) and adult rats respectively.
Methods:
The neural stem cells were isolated from the hippocampi of neonatal and adult rats and cultured in serum-free medium containing basic fibroblast growth factor (bFGF), brain-derived neurotrophic factor (BDNF) and other growth factors using single-cell cloning technique. Immunochemistry was employed to identify the cloned cells and their daughter cells as well as mature neural cells differentiated from the cell clone.
Results:
The floating neural spheres obtained by above culture procedures for isolated cells from the hippocampi of neonatal and adult rats were positive for nestin expression, with the potential for further cloning and capable of differentiation into neurons or glial cells, which formed reticular connection between the cells and expressed neuron-specific enolase (NSE) and glial fibrillary acidic protein (GFAP) respectively.
Conclusion:
The hippocampi of neonatal and adult rats contain neural stem cells.
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