Journal
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
Volume 108, Issue 17, Pages 6993-6998Publisher
NATL ACAD SCIENCES
DOI: 10.1073/pnas.1102559108
Keywords
3D cell culture; cellular mechanics; systems biology; multicellular aggregate
Categories
Funding
- Materials Research Science and Engineering Centers of the National Science Foundation [DMR-0520651]
- Nanotechnology Interdisciplinary Research Team [DMI-0506661]
- National Institutes of Health [R01EB008664-01A1]
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In a nonadhesive environment, cells will self-assemble into microtissues, a process relevant to tissue engineering. Although this has been recognized for some time, there is no basis for quantitative characterization of this complex process. Here we describe a recently developed assay designed to quantify aspects of the process and discuss its application in comparing behaviors between cell types. Cells were seeded in nonadhesive micromolded wells, each well with a circular trough at its base formed by the cylindrical sidewalls and by a central peg in the form of a right circular cone. Cells settled into the trough and coalesced into a toroid, which was then driven up the conical peg by the forces of self-assembly. The mass of the toroid and its rate of upward movement were used to calculate the cell power expended in the process against gravity. The power of the toroid was found to be 0.31 +/- 0.01 pJ/h and 4.3 +/- 1.7 pJ/h for hepatocyte cells and fibroblasts, respectively. Blocking Rho kinase by means of Y-27632 resulted in a 50% and greater reduction in power expended by each type of toroid, indicating that cytoskeletal-mediated contraction plays a significant role in the self-assembly of both cell types. Whereas the driving force for self-assembly has often been viewed as the binding of surface proteins, these data show that cellular contraction is important for cell-cell adhesion. The power measurement quantifies the contribution of cell contraction, and will be useful for understanding the concerted action of the mechanisms that drive self-assembly.
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