Journal of Applied Physiology, Vol 43, Issue 5 902-906, Copyright © 1977 by American Physiological Society
Metabolism in rat lung tissue slices: technical factors
J. J. O'Neil, R. L. Sanford, S. Wasserman and D. F. Tierney
We compared several sets of conditions used to estimate metabolism in rat
lung slices. 14CO2 production from [14C]glucose, oxygen consumption,
lactate production, and glucose consumption were used as measures of
metabolic activity. The calculated results differed when we used 1)
different techniques for estimating tissue weight, 2) tissue slices of
0.3-, 0.5-, 0.7-, and 1.0-mm thickness, 3) 95% air or 95% oxygen with 5%
CO2 4) a delay after slice preparation and 4 degrees C and room temperature
or periods of anoxia before incubation, 5) shaking rates of 60, 90, 120, or
150/min, 6) phosphate or bicarbonate buffers. Conditions of maximal
activity were found using 95% O2 with 1.0-mm tissue slices, shaking at
120/min in phosphate buffer without periods of hypoxia or undue delays
before incubation. Tissue weight should be obtained without exposure to
aqueous solutions or dehydration by contact with cotton gauze or filter
paper.
