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细胞拉伸器:一种用于对细胞群体进行机械刺激的新型装置。

The cell-stretcher: A novel device for the mechanical stimulation of cell populations.

作者信息

Seriani S, Del Favero G, Mahaffey J, Marko D, Gallina P, Long C S, Mestroni L, Sbaizero O

机构信息

Department of Engineering and Architecture, University of Trieste, Trieste, Italy.

Department of Food Chemistry and Toxicology, University of Vienna, Vienna, Austria.

出版信息

Rev Sci Instrum. 2016 Aug;87(8):084301. doi: 10.1063/1.4959884.

Abstract

Mechanical stimulation appears to be a critical modulator for many aspects of biology, both of living tissue and cells. The cell-stretcher, a novel device for the mechanical uniaxial stimulation of populations of cells, is described. The system is based on a variable stroke cam-lever-tappet mechanism which allows the delivery of cyclic stimuli with frequencies of up to 10 Hz and deformation between 1% and 20%. The kinematics is presented and a simulation of the dynamics of the system is shown, in order to compute the contact forces in the mechanism. The cells, following cultivation and preparation, are plated on an ad hoc polydimethylsiloxane membrane which is then loaded on the clamps of the cell-stretcher via force-adjustable magnetic couplings. In order to show the viability of the experimentation and biocompatibility of the cell-stretcher, a set of two in vitro tests were performed. Human epithelial carcinoma cell line A431 and Adult Mouse Ventricular Fibroblasts (AMVFs) from a dual reporter mouse were subject to 0.5 Hz, 24 h cyclic stretching at 15% strain, and to 48 h stimulation at 0.5 Hz and 15% strain, respectively. Visual analysis was performed on A431, showing definite morphological changes in the form of cellular extroflections in the direction of stimulation compared to an unstimulated control. A cytometric analysis was performed on the AMVF population. Results show a post-stimulation live-dead ratio deviance of less than 6% compared to control, which proves that the environment created by the cell-stretcher is suitable for in vitro experimentation.

摘要

机械刺激似乎是生物许多方面的关键调节因素,包括活组织和细胞。本文描述了一种细胞拉伸器,这是一种用于对细胞群体进行机械单轴刺激的新型装置。该系统基于可变行程凸轮 - 杠杆 - 挺杆机构,可提供频率高达10Hz、形变量在1%至20%之间的循环刺激。文中给出了运动学分析,并展示了系统动力学的模拟,以计算机构中的接触力。细胞经过培养和制备后,接种在特制的聚二甲基硅氧烷膜上,然后通过力可调磁耦合加载到细胞拉伸器的夹具上。为了证明细胞拉伸器实验的可行性和生物相容性,进行了一组两项体外测试。来自双报告基因小鼠的人上皮癌细胞系A431和成体小鼠心室成纤维细胞(AMVF)分别接受了0.5Hz、15%应变下24小时的循环拉伸,以及0.5Hz、15%应变下48小时的刺激。对A431进行了视觉分析,结果显示与未刺激的对照相比,在刺激方向上出现了明显的细胞外翻形式的形态变化。对AMVF群体进行了细胞计数分析。结果表明,与对照相比,刺激后的活死比偏差小于6%,这证明细胞拉伸器所营造的环境适用于体外实验。

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