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Am J Physiol Cell Physiol 274: C1283-C1289, 1998;
0363-6143/98 $5.00
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Vol. 274, Issue 5, C1283-C1289, May 1998

Is cytoskeletal tension a major determinant of cell deformability in adherent endothelial cells?

Jacob Pourati1, Andrew Maniotis2, David Spiegel1, Jonathan L. Schaffer3, James P. Butler1, Jeffrey J. Fredberg1, Donald E. Ingber2, Dimitrijie Stamenovic4, and Ning Wang1

1 Physiology Program, Harvard School of Public Health and 2 Departments of Pathology and Surgery, Children's Hospital and Harvard Medical School, Boston 02115; and 3 Departments of Orthopedic Surgery, Brigham and Women's Hospital, Children's Hospital, and Harvard Medical School and 4 Department of Biomedical Engineering, Boston University, Boston, Massachusetts 02215

We tested the hypothesis that mechanical tension in the cytoskeleton (CSK) is a major determinant of cell deformability. To confirm that tension was present in adherent endothelial cells, we either cut or detached them from their basal surface by a microneedle. After cutting or detachment, the cells rapidly retracted. This retraction was prevented, however, if the CSK actin lattice was disrupted by cytochalasin D (Cyto D). These results confirmed that there was preexisting CSK tension in these cells and that the actin lattice was a primary stress-bearing component of the CSK. Second, to determine the extent to which that preexisting CSK tension could alter cell deformability, we developed a stretchable cell culture membrane system to impose a rapid mechanical distension (and presumably a rapid increase in CSK tension) on adherent endothelial cells. Altered cell deformability was quantitated as the shear stiffness measured by magnetic twisting cytometry. When membrane strain increased 2.5 or 5%, the cell stiffness increased 15 and 30%, respectively. Disruption of actin lattice with Cyto D abolished this stretch-induced increase in stiffness, demonstrating that the increased stiffness depended on the integrity of the actin CSK. Permeabilizing the cells with saponin and washing away ATP and Ca2+ did not inhibit the stretch-induced stiffening of the cell. These results suggest that the stretch-induced stiffening was primarily due to the direct mechanical changes in the forces distending the CSK but not to ATP- or Ca2+-dependent processes. Taken together, these results suggest preexisting CSK tension is a major determinant of cell deformability in adherent endothelial cells.

mechanical tension; shape stability; cell adhesion; shear deformation; stiffness


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