Influence of glutaraldehyde fixation of cells adherent to solid substrata on their detachment during exposure to shear stress |
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Authors: | T. G. van Kooten J. M. Schakenraad H. C. van der Mei H. J. Busscher |
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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 |
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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. |
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Keywords: | Glutaraldehyde fixation detachment human fibroblasts parallel plate flow chamber |
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