A disk-type apparatus for applying fluid shear stress on cultured endothelial cell

Abstract

To study the effect of fluid shear stress on cultured endothelial cells, we have developed an apparatus for the stress creation, which consists of a stainless steel disk driven by an electric DC motor and a stage to place a culture dish and to adjust the distance between the disk and the dish. When the disk is rotated, a concentric fluid movement occurs in the culture medium in the dish and exerts the shear stress on the endothelial cells cultured on the bottom of the dish. A theoretical analyses concerning the induced concentric flow velocity predicted that when the angular velocity of the disk rotation (ω) is slow enough to maintain a Reynolds’ number of the order of 10, the exerted wall shear stress ${\mathit{\tau }}_{\mathrm{w}}$ on the endothelial cell monolayer is given for a constant as ${\mathit{\tau }}_{\mathrm{w}}=\mathit{\mu }\mathrm{r}\mathit{\omega }\mathrm{/}\mathrm{d}$ where μ is the viscosity of the medium, d the distance from the plate to the monolayer and r the radial distance from the center of the dish. When ω is varied in a sinusoidal mode ${\mathit{\tau }}_{\mathrm{w}}$ also becomes sinusoidal, thus allowing to apply a pulsatile stress. In vitro experiments carried out to examine the validity of the theoretical results, using a suspension of polystyrene as a tracer with the ordinary culture medium and 99% ethanol, revealed excellent agreement of the measured velocity profiles with the predicted ones. The results demonstrated that the present apparatus can create both the steady and pulsatile wall shear stress on the culture cell layer as expected, unless Reynolds’ number greatly exceeds the level of 10.