A SIMPLE TECHNIQUE TO EFFICIENTLY DISSOCIATE PRIMARY AUDITORY NEURONSFROM 5-DAY-OLD RAT COCHLEAS

The aim of this work was to develop a simple and reproducible method o f dissociation of cochlear spiral ganglion neurons in the rat. This te chnique, wich was developped in 5 day-old rat pups, was based on the u se of a single enzyme, thermolysin. It is easy to set up and allows th e collection of a large amount of neurons. These isolated neurons were kept in a definite, serum free culture medium up to 7 days. Neurons w ere 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 indica ted that all isolated cells were healthy even after 7 days in vitro. T he dissociation and culture methods were found very satisfactory and c an be easily adapted to any kind of experiment requiring isolated spir al ganglion neurons. (C) 1997 Elsevier Science B.V.