Laboratory for Pain Research, University of Split School of Medicine, Split 21000, Croatia.
Pain Center, Department of Anesthesiology, Lausanne University Hospital and Department of Fundamental Neurosciences, University of Lausanne, Lausanne 1011, Switzerland.
J Neurosci Methods. 2024 Jul;407:110143. doi: 10.1016/j.jneumeth.2024.110143. Epub 2024 Apr 25.
Silicon-based micro-pillar substrates (MPS), as three-dimensional cell culture platforms with vertically aligned micro-patterned scaffolding structures, are known to facilitate high-quality growth and morphology of dorsal root ganglion (DRG) sensory neurons, promote neurite outgrowth and enhance neurite alignment. However, the electrophysiological aspects of DRG neurons cultured on silicon MPSs have not been thoroughly investigated, which is of greatest importance to ensure that such substrates do not disrupt neuronal homeostasis and function before their widespread adoption in diverse biomedical applications.
We conducted whole-cell patch-clamp recordings to explore the electrophysiological properties of DRG neurons cultured on MPS arrays, utilizing a custom-made upright patch-clamp setup.
Our findings revealed that DRG neurons exhibited similar electrophysiological responses on patterned MPS samples when compared to the control planar glass surfaces. Notably, there were no significant differences observed in the action potential parameters or firing patterns of action potentials between neurons grown on either substrate.
In the current study we for the first time confirmed that successful electrophysiological recordings can be obtained from the cells grown on MPS.
Our results imply that, despite the potential alterations caused by the cumulative trauma of tissue harvest and cell dissociation, essential functional cell properties of DRG neurons appear to be relatively maintained on MPS surfaces. Therefore, vertically aligned silicon MPSs could be considered as a potentially effective three-dimensional system for supporting a controlled cellular environment in culture.
硅基微柱基底(MPS)作为具有垂直排列微图案支架结构的三维细胞培养平台,已知其有利于背根神经节(DRG)感觉神经元的高质量生长和形态,促进神经突生长并增强神经突取向。然而,在广泛应用于各种生物医学应用之前,尚未彻底研究在硅 MPS 上培养的 DRG 神经元的电生理方面,这对于确保此类基底不破坏神经元内稳态和功能至关重要。
我们使用定制的直立式膜片钳装置进行全细胞膜片钳记录,以研究 MPS 阵列上培养的 DRG 神经元的电生理特性。
我们的研究结果表明,与对照平面玻璃表面相比,在图案化 MPS 样品上培养的 DRG 神经元表现出相似的电生理反应。值得注意的是,在两种底物上生长的神经元的动作电位参数或动作电位发射模式均无明显差异。
在本研究中,我们首次证实可以从生长在 MPS 上的细胞中获得成功的电生理记录。
我们的结果表明,尽管组织收获和细胞解离的累积创伤可能会导致潜在的改变,但 DRG 神经元的基本功能细胞特性似乎在 MPS 表面相对保持。因此,垂直排列的硅 MPS 可以被认为是支持受控细胞环境的有效三维系统。