M. Brehm et al., QUANTIFICATION OF REACTIVE OXYGEN SPECIES GENERATED BY ALVEOLAR MACROPHAGES USING LUCIGENIN-ENHANCED CHEMILUMINESCENCE - METHODICAL ASPECTS, Toxicology letters, 87(2-3), 1996, pp. 131-138
Alveolar macrophages lavaged from bovine lungs using Ca2+- and Mg2+-de
prived saline containing EGTA for calcium chelation were cultivated in
RPMI-1640 medium. The generation of reactive oxygen species (ROS) was
assessed by determination of lucigenin-enhanced chemiluminescence (LU
C-CL) using a LB 9505 C Biolumat. Monitoring of LUC-CL depends on vari
ous methodical parameters: besides medium constituents, the method of
cell harvesting and the time-schedule of the protocol appear to be ess
ential parameters that influence ROS-generation. In addition, the cell
number (cell density) as well as the ratio of cell number to particle
mass influence the amount of ROS-generation. Following exposure of ce
lls to micronized quartz, we observed a dose-related increase in the g
eneration of ROS.