[The isolating culture of neural stem cells from rat's fetus brain and transfection by electroporation].
OBJECTIVE
To isolate and culture the neural stem cells (NSC) from rat's fetus brain, to study the transfection efficacy of NSC using electroporation.


METHODS
We isolated, cultured and amplified NSC from the brain of SD fetal rats. NSC and differentiated cells were identified using immunofluorescent histochemical methods. Using green fluorescence protein (GFP) as the marker, pEGFP-N1 was transfected into NSC using electroporation, and the transfection rate was calculated by counting the NSCs with green fluorescent.


RESULTS
The cells isolated from brain tissue of fetal rats can proliferate for long time, both primary and passage culture of NSC can express specific antigen of NSC-Nestin, and after induced differentiation, the cells can express specific antigen of astrocytes-Glial fibrillary acidic protein (GFAP) and specific antigen of neurons-Neuron-specific enolase (NSE). The transfection rates of electroporation were 17.9% - 69.1%, the average was 30.5%.


CONCLUSIONS
Isolated NSC which had the features of self-proliferation and differentiation potential from brain tissue of SD fetal rats, confirmed that electroporation was an efficient transfection method for NSC, provided experimental basis for gene therapy in the future.
Liu
Chun-jiang
CJ
Department of Pediatrics, Xiangya Hospital, Central South University, Changsha, China.
Yin
Fei
F
Zheng
Xiang-rong
XR
Zhang
Shan-shan
SS
Tan
Jie-lu
JL
D000818:Animals; D001921:Brain / Q000166:cytology* / Q000196:embryology; D002454:Cell Differentiation; D002478:Cells, Cultured; D018274:Electroporation; D009474:Neurons / Q000166:cytology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D014162:Transfection