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PROTEIN AND VESICLE TRAFFICKING, CYTOSKELETON

Single membrane tether extraction from adult and neonatal dermal microvascular endothelial cells

Department of Biomedical Engineering, Washington University, Saint Louis, Missouri

Submitted 8 May 2006 ; accepted in final form 24 October 2006

Membrane tethers were found to be extracted from leukocytes and macrovascular endothelial cells (e.g., human umbilical vein endothelial cells or HUVECs) when a point pulling force was exerted. These tethers stabilize leukocyte rolling on the endothelium during the inflammatory response. However, little is known about tether extraction from other vascular cells like microvascular endothelial cells (MECs). In this study, we extracted tethers from both adult and neonatal dermal MECs with the micropipette aspiration technique. We found a linear relationship between the pulling force and tether growth velocity for both cell lines. This constitutive relationship is mainly determined by the membrane mechanical property and the underlying actin-based cytoskeleton for both attached and suspended endothelial cells. It is independent of cell surface receptor type, attachment state, cytokine stimulation, or cell lineage. For both types of MECs, the threshold forces are 50 pN and the effective viscosities are around 0.5 pN·s/µm. These results, which are close to what was obtained from HUVECs, indicate that homogeneity is preserved in terms of tether extraction among different types of endothelial cells, and simultaneous tethers are likely extracted when leukocytes roll on either microvascular or macrovascular surfaces.

leukocyte rolling; cell mechanics; micropipette; cytoskeleton