Measurement of Aggregate Cohesion by Tissue Surface Tensiometry
Measurement of Aggregate Cohesion by Tissue Surface Tensiometry
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DOI：
10.3791/2739
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发表时间：
2011-04-01
影响因子：
1.2
通讯作者：
Foty, Ramsey A.
中科院分区：
文献类型：
--
作者：
Butler, Christine M.;Foty, Ramsey A.
细胞间结合能的严格测量只能使用基于热力学原理的方法在平衡系统中进行。我们开发了组织表面张力测量仪(TST)，专门用来测量细胞间相互作用的表面自由能。先前已详细描述了作为TST基础的生物物理概念(1，2)。该方法的基础是观察到，如果在摇床培养中保持相互粘连的细胞，会自发地组装成簇。随着时间的推移，这些星团会聚集在一起形成球体。这种四舍五入的行为模仿了液体系统的行为特征。细胞间结合能是通过在定制的组织表面张力计中压缩平行平板之间的球形聚集体来测量的。用来测量液滴表面张力的同一数学方程也被用来测量3D组织状球形聚集体的表面张力。液体表面张力的细胞等价物是细胞间结合能，或者更广泛地说，是组织凝聚力。我们实验室以前的研究表明，组织表面张力(1)预测两组胚胎细胞将如何相互作用(1-5)，(2)可以强烈影响组织与生物材料相互作用的能力(6)，(3)不仅可以通过直接操纵基于钙粘附素的细胞间凝聚力(7)，而且可以通过操纵关键的ECM分子如FN8-11和4)与肺癌(12)、纤维肉瘤(13)、脑瘤(14)和前列腺肿瘤细胞系(15)的侵袭潜力相关。在本文中，我们将描述该设备，详细说明生成球体、将球体加载到张力计腔体、启动聚合压缩以及分析和验证生成的组织表面张力测量所需的步骤。
Rigorous measurement of intercellular binding energy can only be made using methods grounded in thermodynamic principles in systems at equilibrium. We have developed tissue surface tensiometry (TST) specifically to measure the surface free energy of interaction between cells. The biophysical concepts underlying TST have been previously described in detail(1,2). The method is based on the observation that mutually cohesive cells, if maintained in shaking culture, will spontaneously assemble into clusters. Over time, these clusters will round up to form spheres. This rounding-up behavior mimics the behavior characteristic of liquid systems. Intercellular binding energy is measured by compressing spherical aggregates between parallel plates in a custom-designed tissue surface tensiometer. The same mathematical equation used to measure the surface tension of a liquid droplet is used to measure surface tension of 3D tissue-like spherical aggregates. The cellular equivalent of liquid surface tension is intercellular binding energy, or more generally, tissue cohesivity. Previous studies from our laboratory have shown that tissue surface tension (1) predicts how two groups of embryonic cells will interact with one another(1-5), (2) can strongly influence the ability of tissues to interact with biomaterials(6), (3) can be altered not only through direct manipulation of cadherin-based intercellular cohesion(7), but also by manipulation of key ECM molecules such as FN8-11 and 4) correlates with invasive potential of lung cancer(12), fibrosarcoma(13), brain tumor(14) and prostate tumor cell lines(15). In this article we will describe the apparatus, detail the steps required to generate spheroids, to load the spheroids into the tensiometer chamber, to initiate aggregate compression, and to analyze and validate the tissue surface tension measurements generated.