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A novel collagen gel-based measurement technique for quantitation of cell contraction force
Authors:Tianrong Jin  Li Li  Richard C M Siow  Kuo-Kang Liu
Affiliation:1.School of Engineering, University of Warwick, Coventry CV4 7AL, UK;2.Cardiovascular Division, British Heart Foundation Centre of Research Excellence, Faculty of Life Sciences and Medicine, King''s College London, London SE1 9NH, UK
Abstract: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 µM) was used to stimulate fibroblast contraction while the myosin light chain kinase inhibitor ML-7 (25 µM) 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.
Keywords:collagen gel  human aortic adventitial fibroblast  histamine  cell contraction force  nano-indentation  elasticity
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