A simple technique to efficiently dissociate primary auditory neurons from 5 day-old rat cochleas.
The aim of this work was to develop a simple and reproducible method of dissociation of cochlear spiral ganglion neurons in the rat. This technique, wich was developed in 5 day-old rat pups, was based on the use of a single enzyme, thermolysin. It is easy to set up and allows the collection of a large amount of neurons. These isolated neurons were kept in a definite, serum free culture medium up to 7 days. Neurons were characterized both by standard morphological criteria and by using a specific neuronal marker (anti-neurofilament 200 kD) after 2 h and 7 days in culture. Cell viability, assessed by fluorescent dyes indicated that all isolated cells were healthy even after 7 days in vitro. The dissociation and culture methods were found very satisfactory and can be easily adapted to any kind of experiment requiring isolated spiral ganglion neurons.
Ripoll
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INSERM U. 254, and Université de Montpellier I, Neurobiologie de l'Audition, Plasticité Synaptique, CHR Saint Charles, France. biomol@soleil.mnet.fr
Rebillard
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G
D000818:Animals; D002452:Cell Count; D002469:Cell Separation / Q000379:methods*; D002470:Cell Survival; D002478:Cells, Cultured; D003051:Cochlea / Q000166:cytology*; D007150:Immunohistochemistry; D009474:Neurons / Q000166:cytology* / Q000378:metabolism; D009488:Neurosciences / Q000379:methods*; D051381:Rats; D013136:Spiral Ganglion / Q000166:cytology* / Q000378:metabolism