[Isolation, culture and identification of rat hippocampal neural stem cells].
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.


CONCLUSIONS
The hippocampi of neonatal and adult rats contain neural stem cells.
Department of High-temperature Medicine, Southern Medical University, Guangzhou 510515, China.
Hong-bo
HB
Guo
Fei
F
Zou
D000818:Animals; D000831:Animals, Newborn; D019208:Brain-Derived Neurotrophic Factor / Q000494:pharmacology; D002454:Cell Differentiation / Q000502:physiology*; D002469:Cell Separation; D002478:Cells, Cultured; D005260:Female; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D005904:Glial Fibrillary Acidic Protein / Q000096:biosynthesis; D006624:Hippocampus / Q000166:cytology*; D008297:Male; D009474:Neurons / Q000166:cytology*; D010751:Phosphopyruvate Hydratase / Q000096:biosynthesis; D051381:Rats; D017207:Rats, Sprague-Dawley; D013234:Stem Cells / Q000166:cytology*
D019208:Brain-Derived Neurotrophic Factor; D005904:Glial Fibrillary Acidic Protein; D016222:Fibroblast Growth Factor 2; D010751:Phosphopyruvate Hydratase