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Membrane Tether Extraction from Human Umbilical Vein Endothelial Cells and Its Implication in Leukocyte Rolling

Department of Biomedical Engineering, Washington University, St. Louis, Missouri 63130

Correspondence: Address reprint requests to Jin-Yu Shao, PhD, Dept. of Biomedical Engineering, Washington University in St. Louis, Campus Box 1097, Rm. 290E Whitaker Hall, One Brookings Dr., St. Louis, MO 63130-4899. Tel.: 314-935-7467; Fax: 314-935-7448; E-mail: shao{at}biomed.wustl.edu.

During the rolling of human neutrophils on the endothelium, tethers (cylindrical membrane tubes) are likely extracted from the neutrophil. Tether extraction reduces the force imposed on the adhesive bond between the neutrophil and endothelium, thereby facilitating the rolling. However, whether tethers can be extracted from the endothelium is still unknown. Here, with the micropipette-aspiration technique, we show that tethers can be extracted from either suspended or attached human umbilical vein endothelial cells. We also show that a linear relationship between the pulling force and tether growth velocity exists and this relationship does not depend on the receptor type (used to impose point forces), tumor necrosis factor- stimulation, or cell attachment state. With linear regression, we determined that the threshold force was 50 pN and the effective viscosity was 0.50 pN·s/µm. Therefore, tethers might be simultaneously extracted from the neutrophil and endothelial cell during the rolling and, more importantly, the endothelial cell might contribute much more to the total composite tether length than the neutrophil. Compared with tether extraction from the neutrophil alone, simultaneous tether extraction results in a larger increase in the lifetime of the adhesive bond, and thus further stabilizes the rolling of neutrophils under high physiological shear stresses.

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