基于明胶的柔性三维毛细管模式微加工技术在细胞集体迁移分析中的应用

Hiromichi Hashimoto, Mitsuru Sentoku, Kento Iida, K. Yasuda
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引用次数: 0

摘要

细胞集体迁移被认为是细胞群的动态和相互作用行为,是生物体多种生理发育所必需的。最近的研究表明,环境的地形特性调节着细胞群的迁移模式,如扩散与收缩弛豫运输以及在更大的可用空间中出现的漩涡。然而,常规的体外实验无法观察到细胞行为随结构变化的变化。在这里,我们开发了一种方法来制造毛细血管微隧道的柔性三维结构来检测血管内皮细胞(ECs)的行为。通过聚焦1064 nm可渗透红外激光照射微针尖端µm大小的吸收区,对部分明胶进行点加热,在明胶内部形成直径变化的微隧道。内皮细胞在毛细血管微隧道内表面呈单层二维移动和扩散,而不是填充毛细血管。与3D直线地形约束表现出宽度依赖的迁移速度不同,前导ECs的迁移速度随着前导ECs后面的细胞供应的变化而变化,这是由直径改变结构的进展引起的。我们的发现为三维约束结构中的集体迁移特性提供了见解,这些特性具有细胞数量守恒的流体样行为。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Development of Gelatin-Based Flexible Three-Dimensional Capillary Pattern Microfabrication Technology for Analysis of Collective Cell Migration
The collective cell migration is thought to be a dynamic and interactive behavior of cell cohorts which is essential for diverse physiological developments in living organisms. Recent studies revealed that topographical properties of the environment regulate the migration modes of cell cohorts, such as diffusion versus contraction relaxation transport and the appearance of vortices in larger available space. However, conventional in vitro assays fail to observe the change in cells behavior in response to the structural changes. Here, we have developed a method to fabricate the flexible three-dimensional structures of capillary microtunnels to examine the behavior of vascular endothelial cells (ECs). The microtunnels with altering diameters were formed inside gelatin-gel by spot heating a portion of gelatin by irradiating the µm-sized absorption at the tip of the microneedle with a focused permeable 1064 nm infrared laser. The ECs moved and spread two-dimensionally on the inner surface of capillary microtunnels as monolayer instead of filling the capillary. In contrast to the 3D straight topographical constraint, which exhibited width dependence migration velocity, leading ECs altered its migration velocity accordingly to the change in supply of the cells behind the leading ECs, caused by the progression through the diameter altering structure. Our findings provide insights into the collective migration properties in 3D confinement structures as fluid-like behavior with conservation of cell numbers.
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