Jin Tianrong, Li Li, Siow Richard C M, Liu Kuo-Kang
J R Soc Interface. 2015 May 6;12(106). doi: 10.1098/rsif.2014.1365.
Cell contraction force plays an important role in wound healing, inflammation,angiogenesis and metastasis. This study describes a novel method to quantify single cell contraction force in vitro using human aortic adventitial fibroblasts embedded in a collagen gel. The technique is based on a depth sensing nano-indentation tester to measure the thickness and elasticity of collagen gels containing stimulated fibroblasts and a microscopy imaging system to estimate the gel area. In parallel, a simple theoretical model has been developed to calculate cell contraction force based on the measured parameters. Histamine (100 mM) was used to stimulate fibroblast contraction while the myosin light chain kinase inhibitor ML-7 (25 mM) was used to inhibit cell contraction. The collagen matrix used in the model provides a physiological environment for fibroblast contraction studies. Measurement of changes in collagen gel elasticity and thickness arising from histamine treatments provides a novel convenient technique to measure cell contraction force within a collagen matrix. This study demonstrates that histamine can elicit a significant increase in contraction force of fibroblasts embedded in collagen,while the Young's modulus of the gel decreases due to the gel degradation.
细胞收缩力在伤口愈合、炎症、血管生成和转移中起着重要作用。本研究描述了一种使用包埋在胶原凝胶中的人主动脉外膜成纤维细胞在体外定量单细胞收缩力的新方法。该技术基于一个深度传感纳米压痕测试仪来测量含有受刺激成纤维细胞的胶原凝胶的厚度和弹性,以及一个显微镜成像系统来估计凝胶面积。同时,已开发出一个简单的理论模型,根据测量参数计算细胞收缩力。使用组胺(100 mM)刺激成纤维细胞收缩,而使用肌球蛋白轻链激酶抑制剂ML-7(25 mM)抑制细胞收缩。模型中使用的胶原基质为成纤维细胞收缩研究提供了一个生理环境。测量组胺处理引起的胶原凝胶弹性和厚度变化,提供了一种在胶原基质内测量细胞收缩力的新颖便捷技术。本研究表明,组胺可显著增加包埋在胶原中的成纤维细胞的收缩力,而凝胶的杨氏模量由于凝胶降解而降低。