期刊
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS
卷 -, 期 50, 页码 -出版社
JOURNAL OF VISUALIZED EXPERIMENTS
DOI: 10.3791/2739
关键词
Bioengineering; Issue 50; 3D; aggregate cohesion; tissue surface tension; parallel plate compression
资金
- NCI NIH HHS [R01 CA118755] Funding Source: Medline
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.
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