Primary culture of rat hippocampal neurons and detection of the neuronal excitability / 南方医科大学学报
Journal of Southern Medical University
;
(12): 2080-2083, 2010.
Article
Dans Chinois
| WPRIM
| ID: wpr-330778
ABSTRACT
<p><b>OBJECTIVE</b>To improve the efficiency of primary culture of hippocampal neurons and obtain highly purified neurons with good in vitro growth and minimal risk of contamination.</p><p><b>METHODS</b>The hippocampal neurons of neonatal Wistar rats were isolated and the single cell suspension was prepared by mechanical trituration and sedimentation in stead of trypsin digestion and filteration. Twenty-four hours after the cell plating, the culture medium was removed and replaced by serum-free DMEM/F12 with B27 supplementation. Half of the culture medium was changed 2-3 times every week. The morphological changes of the neurons were observed under inverted phase-contrast microscope. Immunofluorescence staining for NSE was performed to identify the neurons, and the purity of neurons was calculated. The hippocampal neurons were stained with calcium-sensitive fluorescent dye to monitor the effect of KCl on neuronal excitability by a calcium imaging system.</p><p><b>RESULTS AND CONCLUSION</b>This simplified method is time-saving and cost-effective for primary culture of hippocampal neurons with reduced risk of contamination, and the neurons obtained showed high uniformity, purity and long-term viability.</p>
Texte intégral:
Disponible
Indice:
WPRIM (Pacifique occidental)
Sujet Principal:
Physiologie
/
Potentiels d'action
/
Milieux de culture sans sérum
/
Rat Wistar
/
Biologie cellulaire
/
Culture de cellules primaires
/
Hippocampe
/
Animaux nouveau-nés
/
Méthodes
/
Neurones
Type d'étude:
Etude diagnostique
Limites du sujet:
Animaux
langue:
Chinois
Texte intégral:
Journal of Southern Medical University
Année:
2010
Type:
Article
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