Goldenberg S S, De Boni U
J Neurobiol. 1983 May;14(3):195-206. doi: 10.1002/neu.480140304.
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.
非神经元细胞使体外神经元的研究变得复杂。使用Percoll梯度可以轻松获得纯的活神经元群体。对于每个实验，将20个背根神经节（DRG）切碎，然后依次用胶原酶和胰蛋白酶解离，这些酶会消化所有细胞间连接。首先根据密度分离解离后的组织，产生富含神经元和卫星细胞的中间相部分，然后再根据大小进一步分离。最终沉淀中获得的神经元数量约为50,000个（每个DRG 2500个），通常最初的活力超过80%，纯度超过90%。通过培养中神经突生长的出现来确认神经元的活力。因此，通过一种简单快速的方法可以获得纯的且有活力的神经元群体。
