螺旋神经节神经元(SGN)的体外培养是研究神经元行为的许多方面并确定SGN保护和再生的潜在治疗靶标的有用方法。然而,SGN的分离以及在体外长期维持其结构和功能仍然具有挑战性.在这项研究中,我们通过荧光激活细胞分选从Bhlhb5-cre和Rosa26-tdTomato小鼠中分离SGN,并确定细胞纯度。然后我们将纯SGN封装在基质胶中,并在体外培养SGN。我们发现,三维(3D)-matrigel培养环境显著抑制凋亡和提高SGN的存活,这使得SGN的长期培养长达6个月。3D-matrigel系统还显著促进了SGN的神经突生长,增加了细胞的极性,促进了生长锥的面积,并显着增加了SGN的突触密度。更重要的是,3D-matrigel系统有助于维持和促进SGN的电生理特性。总之,3D-matrigel培养系统促进了纯化SGN的体外存活,并保持了其形态结构和功能,因此可以成为研究长期培养中纯化SGN的生理和病理生理学的有用工具。
In vitro culture of spiral ganglion neurons (SGNs) is a useful approach to investigate numerous aspects of neuronal behavior and to identify potential therapeutic targets for SGN protection and regeneration. However, the isolation of SGNs and the long-term maintenance of their structure and function in vitro remain challenging. In this study, we isolated SGNs from Bhlhb5-cre and Rosa26-tdTomato mice with fluorescence-activated cell sorting and determined the cell purity. We then encapsulated the pure SGNs in matrigel and cultured the SGNs in vitro. We found that the three-dimensional (3D)-matrigel culture environment significantly suppressed apoptosis and improved SGN survival in vitro, which enabled the long-term culture of SGNs for up to 6 months. The 3D-matrigel system also significantly promoted neurite outgrowth of the SGNs, increased the cells\' polarity, promoted the area of growth cones, and significantly increased the synapse density of the SGNs. More importantly, the 3D-matrigel system helped to maintain and promote the electrophysiological properties of the SGNs. In conclusion, the 3D-matrigel culture system promoted the survival of purified SGNs in vitro and maintained their morphological structure and function and thus could be a useful tool for studying the physiology and pathophysiology of purified SGNs in long-term culture.