Prostaglandin and thromboxane production by rat macrophages (Abstract)

Rat macrophages collected by peritoneal lavage were plated out at a concentration of 2x×107 cells per 25 cm2 in Medium 199 containing 5% fetal calf serum, 25 m mol/1 HEPES pH 7.4 and antibiotics. The cells were aged for 24 h prior to use producing a pure macrophage population (>95%) as shown by the presence of Fc receptor sites, the ability to phagocytose sheep red blood cells and the presence of non-specific esterase staining granules. The cells were cultured for a further 48 h in fresh medium in the presence or absence of serum opsonized zymosan. Following removal of cells by centrifugation media were acidified and extracted with diethyl ether. The prostaglandins and thromboxane B2 in the sample were then derivatized to form their methyl ester, methoxime, tertiary butyldimethylsilyl ethers and the levels of the prostaglandins and thromboxane B2 were measured by quantitative gas chromatography/mass spectrometry.