Quantification of reactive oxygen species generated by alveolar macrophages using lucigenin-enhanced chemiluminescence--methodical aspects.
Alveolar macrophages lavaged from bovine lungs using Ca2(+)- and Mg2(+)-deprived 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 (LUC-CL) using a LB 9505 C Biolumat. Monitoring of LUC-CL depends on various methodical parameters: besides medium constituents, the method of cell harvesting and the time-schedule of the protocol appear to be essential 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 cells to micronized quartz, we observed a dose-related increase in the generation of ROS.
German Cancer Research Center, Department 0420, Heidelberg, Federal Republic of Germany.
M
M
Brehm
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E
Schiller
W J
WJ
Zeller
D000166:Acridines / Q000737:chemistry* / Q000378:metabolism; D000818:Animals; D002118:Calcium / Q000172:deficiency; D002417:Cattle; D002448:Cell Adhesion / Q000187:drug effects; D002452:Cell Count; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004533:Egtazic Acid / Q000378:metabolism; D008163:Luminescent Measurements; D016676:Macrophages, Alveolar / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D008275:Magnesium Deficiency; D011791:Quartz / Q000633:toxicity*; D017382:Reactive Oxygen Species / Q000378:metabolism*
D000166:Acridines; D017382:Reactive Oxygen Species; D011791:Quartz; C033472:10,10'-dimethyl-9,9'-biacridinium; D004533:Egtazic Acid; D002118:Calcium