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Influence of glutaraldehyde fixation of cells adherent to solid substrata on their detachment during exposure to shear stress
Authors:T. G. van Kooten  J. M. Schakenraad  H. C. van der Mei  H. J. Busscher
Affiliation:(1) Laboratory for Materia Technica, University of Groningen, Antonius Deusinglaan 1, 9713 AV Groningen, The Netherlands;(2) Centre for Biomedical Technology, University of Groningen, Oostersingel 59, Gb 25, FO, 9713 EZ Groningen, The Netherlands
Abstract:In order to determine the response of fixed and nonfixed cells adherent to a solid substratum to shear stress, human fibroblasts were allowed to adhere and spread on either hydrophilic glass or hydrophobic Fluoroethylene-propylene (FEP-Teflon) and fixed with glutaraldehyde. Then, the cells were exposed to an incrementally loaded shear stress in a parallel plate flow chamber up to shear stresses of about 500 dynes/cm2, followed by exposure to a liquid-air interface passage. The cellular detachment was compared with the one of nonfixed cells. In case of fixed cells, 50% of the adhering cells detached from FEP-Teflon at a shear stress of 350 dynes/cm2, whereas 50% of the adhering, nonfixed cells detached already at a shear stress of 20 dynes/cm2. No fixed cells detached from glass for shear stresses up to at least 500 dynes/cm2. More than 50% of the nonfixed cells were detached from glass at a shear stress of 350 dynes/cm2. Furthermore, the shape and morphology of fixed cells did not change during the incrementally loaded flow, in contrast to the ones of nonfixed cells, which clearly rounded up prior to detachment.
Keywords:Glutaraldehyde fixation  detachment  human fibroblasts  parallel plate flow chamber
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