Using cultures of freshly isolated spiral ganglion cells (SGC) is common to investigate the effect of substances on spiral ganglion neurons (SGN) in vitro. As these cultures contain more cell types than just neurons, and it might be beneficial to have cochlear fibroblasts available to further investigate approaches to reduce the growth of fibrous tissue around the electrode array after cochlear implantation, we aimed at the purification of fibroblasts from the spiral ganglion in the current study. Subcultivation of the primary SGC culture removed the neurons from the culture and increased the fibroblast to glial cell ratio in the preparations, which was revealed by staining for vimentin, the S100B-protein, and the 200-kD neurofilament. We performed direct immunolabeling for the Thy1-glycoprotein and the p75NGFR-enabled fluorescence-based cell sorting. This procedure resulted in a cell culture of cochlear fibroblasts with a purity of more than 99%. The received fibroblasts can be subcultivated for up to 10 passages before proliferation rates drop. Additionally, 80% of the cells survived the first attempt of cryopreservation and exhibited a fibroblast-specific morphology. Using the described approach provides a purified preparation of cochlear fibroblasts, which can now be used in vitro for further investigations.

使用新鲜分离的螺旋神经节细胞 (SGC) 培养物来研究物质对螺旋神经节神经元 (SGN) 的影响是很常见的体外. 由于这些培养物不仅包含神经元，还包含更多的细胞类型，并且拥有耳蜗成纤维细胞以进一步研究减少耳蜗植入后电极阵列周围纤维组织生长的方法可能是有益的，我们的目标是从螺旋中纯化成纤维细胞目前研究中的神经节。原代 SGC 培养物的继代培养从培养物中去除了神经元，并增加了制剂中成纤维细胞与神经胶质细胞的比例，这通过波形蛋白、S100B 蛋白和 200-kD 神经丝的染色来揭示。我们对 Thy1-糖蛋白和 p75NGFR 启用的基于荧光的细胞分选进行了直接免疫标记。该程序产生了纯度超过 99% 的耳蜗成纤维细胞的细胞培养物。在增殖率下降之前，接收到的成纤维细胞最多可传代 10 代。此外，80% 的细胞在第一次冷冻保存尝试中存活并表现出成纤维细胞特异性形态。使用所描述的方法提供了耳蜗成纤维细胞的纯化制剂，现在可以使用体外进行进一步调查。