Pure population of viable neurons from rabbit dorsal root ganglia, using gradients of Percoll

Nonneuronal cells complicate the study of neurons in vitro. A pure population of viable neurons can be obtained easily using gradients of Percoll. For each experiment, 20 dorsal root ganglia (DRG) are minced, then sequentially dissociated in collagenase and trypsin, which digest all the intercellular connections. The dissociated tissue is separated first on the basis of density, creating an interphase fraction enriched in neurons and satellite cells, which are then further separated on the basis of size. The neurons, obtained in the final pellet, number approximately 50,000 (2500 per DRG), routinely exhibit a viability of over 80% initially and are of a purity of over 90%. The viability of the neurons is confirmed by the occurrence of neurite outgrowth in culture. Thus, a pure and viable neuronal population is obtained by a simple and rapid method.