Direct measurement of nitric oxide in headspace gas produced by a chicken macrophage cell line in a closed culture system.
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). HD11 chicken macrophages (1-3 x 10(6)/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 micrograms/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 degrees C. The concentration of NO in the gas sample was quantified using a standard gas mixture of NO (2 microliters/L) in nitrogen. Gas samples from L-arginine-supplemented cultures contained NO (0.028-0.066 pl/microliter), 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.
Sung
Y J
YJ
Institute for Comparative and Environmental Toxicology, Cornell University, Ithaca, NY 14853.
Hotchkiss
J H
JH
Austic
R E
RE
Dietert
R R
RR
D000818:Animals; D002460:Cell Line; D002470:Cell Survival; D002645:Chickens; D046508:Culture Techniques / Q000295:instrumentation / Q000379:methods; D005740:Gases / Q000032:analysis; D006863:Hydrogen-Ion Concentration; D008264:Macrophages / Q000166:cytology / Q000378:metabolism*; D009569:Nitric Oxide / Q000032:analysis* / Q000378:metabolism*; D010101:Oxygen Consumption
D005740:Gases; D009569:Nitric Oxide