Development of stretch stimulation device for three-dimensional culture of PC12 cells
Development of stretch stimulation device for three-dimensional culture of PC12 cells
复制标题
PC12细胞三维培养拉伸刺激装置的研制
DOI：
10.1115/imece2018-86643
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发表时间：
2018
期刊：
影响因子：
--
通讯作者：
Eiji Nakamachi
中科院分区：
文献类型：
--
作者：
Madoka Imura;Ryota Sakiyama;Koji Yamamoto;Yusuke Morita;Eiji Nakamachi
本文报道了利用细胞外环境刺激促进神经轴突的伸展。本研究以牵张刺激为研究对象，建立了一个三维细胞培养系统，模拟体内细胞外基质，探讨了牵张刺激促进轴突延伸的基本机制。将包埋在胶原凝胶中的神经样细胞--大鼠嗜铬细胞瘤细胞（PC 12）注入牵张室中。将其置于牵张刺激装置中，可将应变加载到胶原凝胶上。其次，确定了拉伸室的结构，以实现培养区域的均匀应变分布。使用有限元（FE）分析，我们确认了均匀应变分配在培养区域中的2.7 × 3.0 × 0.5 mm的区域中，该区域是观察区域的候选区域。第三，在单轴循环拉伸刺激（4%应变，1Hz）下进行24小时的PC 12细胞轴突延伸。培养96 h后，用多光子激发荧光显微镜（MPM）观察PC 12细胞的三维形态。最后，我们证实了我们的拉伸刺激装置的可用性和轴突延伸的增强效果。
Enhancement of nerve axonal extension by using the extracellular environmental stimulation were reported. In this study, we focused on the stretch stimulation, and developed a 3D cell culture system to mimic thein vivoextracellular matrices and investigated the fundamental mechanism of axonal extension enhancement.Firstly, we fabricated the stretch stimulation device. The rat phenocromocytoma cells (PC12), the nerve-like cells, embedded in the collagen gel were poured into the stretch chamber. It was set in the stretch stimulation device, which could load the strain to the collagen gel. Secondly, we determined the structure of the stretch chamber to implement the uniform strain distribution in the culture region. Using the finite element (FE) analyses, we confirmed that the uniform strain is assigned in a region of 2.7 × 3.0 × 0.5 mm in the culture region, which is the candidate for the observation region. Thirdly, PC12 cells axonal extension under uniaxial cyclic stretch stimulation (4% strain, 1 Hz) of 24 hours was carried out. After 96 hours’ culture, we observed the 3D morphology of PC12 cells by the multiphoton excitation fluorescence microscope (MPM). Finally, we confirmed the availability of our stretch stimulation device and the enhancement effect of axonal extension.