A Three-Dimensional Culture System with Matrigel Promotes Purified Spiral Ganglion Neuron Survival and Function In Vitro
A Three-Dimensional Culture System with Matrigel Promotes Purified Spiral Ganglion Neuron Survival and Function In Vitro
复制标题
含有基质胶的三维培养系统可促进纯化的螺旋神经节神经元体外存活和功能
DOI：
10.1007/s12035-017-0471-0
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发表时间：
2018-03-01
影响因子：
5.1
通讯作者：
Wang, Haibo
中科院分区：
文献类型：
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作者：
Yan, Wenqing;Liu, Wenwen;Wang, Haibo
螺旋神经节神经元(SGN)的体外培养是研究神经元行为的多个方面以及寻找保护和再生SGN的潜在治疗靶点的有用方法。然而，SGN的分离和在体外长期维持其结构和功能仍然是具有挑战性的。在这项研究中，我们用荧光激活细胞分选法从Bhlhb5-cre和rosa26-td番茄小鼠中分离出SGN，并测定了细胞纯度。然后我们将纯化的SGN包裹在Matrigel中，并在体外培养SGN。我们发现，三维(3D)-Matrigel培养环境显著抑制了SGN的凋亡，并提高了SGN的体外存活，使SGN能够长期培养长达6个月。3D-Matrigel系统还显著促进了SGN的突起生长，增加了细胞的极性，增加了生长锥体的面积，并显著增加了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。综上所述，3D-Matrigel培养系统促进了纯化的SGN在体外的存活，并维持了其形态结构和功能，为长期培养纯化的SGN的生理和病理生理研究提供了一种有用的工具。
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.