Characterization of a computerized assay for rapid and easy determination of leukocyte adhesion to endothelial cells

Abstract

We report on a facile and rapid computerized in-vitro assay for the quantification of leukocyte adhesion to endothelial cells under static conditions using bovine polymorphonuclear neutrophils (PMN) or human leukaemic Mono Mac 6 cells (MM6) and bovine aorta endothelial cells (BAEC). Images of leukocytes adherent to BAEC monolayers grown in microtiter plates were obtained by a digital camera attached to a conventional microscope and transferred to the public domain NIH ImageJ program for analysis. Using individually adapted program routines adherent leukocytes are easily discriminated and reproducibly quantified. The results obtained with our assay correspond to previous findings and demonstrate the suitability of the described procedure, which can easily be adapted to further standards as proven by the use of two different leukocyte species. This assay lends itself to the screening of pharmacological substances with different mechanism of action that might act on either leukocytes or endothelial cells. © 2005 Pharmaceutical Society of Japan.