DIRECT MEASUREMENT OF NITRIC-OXIDE IN HEADSPACE GAS PRODUCED BY A CHICKEN MACROPHAGE CELL-LINE IN A CLOSED CULTURE SYSTEM

被引:20
作者
SUNG, YJ
HOTCHKISS, JH
AUSTIC, RE
DIETERT, RR
机构
[1] CORNELL UNIV,INST COMPARAT & ENVIRONM TOXICOL,110 RICE HALL,ITHACA,NY 14853
[2] CORNELL UNIV,DEPT MICROBIOL IMMUNOL & PARASITOL,ITHACA,NY 14853
[3] CORNELL UNIV,INST FOOD SCI,ITHACA,NY 14853
[4] CORNELL UNIV,DEPT ANIM SCI,ITHACA,NY 14853
关键词
D O I
10.1016/0006-291X(92)91154-I
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
A simple and rapid method was applied for direct measurement of nitric oxide (NO) gas produced by cultured macrophages using a modified chemiluminescence detector, the thermal energy analyzer (TEA). HD 11 chicken macrophages (1-3 × 106/ml) were cultured on microcarrier beads (100 mg/ml) in 140 ml air-tight glass jars (5 ml cell suspension per jar) containing 0.5 μg/ml of LPS and different concentrations of l-arginine. Headspace gas was sampled at 24 hours of culture via a rubber septum and directly injected into a TEA with a liquid nitrogen trap set at -130 to -140°C. The concentration of NO in the gas sample was quantified using a standard gas mixture of NO (2 μl/L) in nitrogen. Gas samples from l-arginine-supplemented cultures contained NO (0.028 - 0.066 pl/μl), whereas NO was not detected in samples from controls. These results suggest that chicken macrophages synthesize NO gas in a dose-dependent manner relative to l-arginine concentration. © 1992.
引用
收藏
页码:36 / 42
页数:7
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