Heat conduction microcalorimetry of overall metabolism in rabbit alveolar macrophages in monolayers and in suspensions.

A technique has been developed for studying the overall metabolism in small numbers of cells in monolayers and in suspensions by measuring the heat exchange rate with a thermopile heat-conduction microcalorimeter. The system was tested with alveolar macrophages from rabbits, and heat flux-time curves could be monitored from numbers of cells as low as 0.05 . 10(6). The metabolic activity was found to be unchanged or to decrease very slowly over a period of 20 h. In a conventional cell medium, without the addition of serum, a mean heat flux value of 19.4 (S.D. 3.2) pW. cell-1 (37 degrees C, pH = 7.4) was obtained for alveolar macrophages in monolayers. The corresponding value for cells in the same medium, with the addition of 20% homologous rabbit serum, was 27.0 (S.D. 2.0). We suggest that this calorimetric method can be used as a short-term cytotoxic test for measuring potentially toxic agents in our environment; this test can involve alveolar macrophages of either animal or human origin. In such an assay, the cells should be used in monolayers, and suspensions should be avoided. We found that when alveolar macrophages were used in suspension, the metabolic activity, measured as heat flux, was dependent on cell concentration, even at values as low as 0.2. 10(6) cells per ml.

[1]  J. Chappell The oxidation of citrate, isocitrate and cis-aconitate by isolated mitochondria. , 1964, The Biochemical journal.

[2]  D. M. Weir,et al.  Handbook of experimental immunology , 1967 .

[3]  J. Suurkuusk,et al.  Operation of a multichannel microcalorimetry system in the micro-submicrowatt region: some methodological aspects. , 1989, Journal of Biochemical and Biophysical Methods.

[4]  V. Ferrans,et al.  Chapter 6 The Human Alveolar Macrophage , 1980 .

[5]  I. Wadsö,et al.  Heat production in human blood lymphocytes. A methodological study. , 1984, Scandinavian journal of clinical and laboratory investigation.

[6]  C. Florén,et al.  Microcalorimetric investigation of metabolism in rat hepatocytes cultured on microplates and in cell suspensions. , 1986, Biochimica et biophysica acta.

[7]  David A. Williams,et al.  Changes in Energy Metabolism, Structure and Function in Alveolar Macrophages under Anaerobic Conditions , 1981, British journal of haematology.

[8]  R. Gronostajski,et al.  Protein degradation in 3T3 cells and tumorigenic transformed 3T3 cells , 1984, Journal of cellular physiology.

[9]  S. Thorén A semiautomatic image analyzer for cell counts in monolayers. II. Application to toxicity estimation and comparisons between two cell-viability markers, fluorescein diacetate and lactate dehydrogenase. , 1989, Journal of toxicology and environmental health.

[10]  R. Bois The alveolar macrophage. , 1985, Thorax.

[11]  D. Golde,et al.  The Pulmonary-Alveolar Macrophage , 1979 .

[12]  J. Brain Macrophage damage in relation to the pathogenesis of lung diseases. , 1980, Environmental health perspectives.

[13]  I. Wadsö,et al.  Thermochemical characterization of T-lymphoma cells under non-growing conditions. , 1986, Cytobios.

[14]  I. Wadsö,et al.  Microcalorimetric measurements of heat production in rabbit alveolar macrophages , 1984 .

[15]  I. Wadsö,et al.  Microcalorimetric measurements on tissue cells attached to microcarriers in stirred suspension. , 1986, Journal of biochemical and biophysical methods.

[16]  S. G. Axline,et al.  Enzymatic basis for bioenergetic differences of alveolar versus peritoneal macrophages and enzyme regulation by molecular O2. , 1977, The Journal of clinical investigation.

[17]  L. Romert,et al.  Effects of air pollutants on the oxidative metabolism and phagocytic capacity of pulmonary alveolar macrophages. , 1983, Journal of toxicology and environmental health.

[18]  J. Lanke,et al.  A semiautomatic image analyzer for cell counts in monolayers. I. Construction, experimental procedure, and precision obtained. , 1989, Journal of toxicology and environmental health.

[19]  P. W. Alberti,et al.  Industrial Noise Pollution and Hearing Impairment. , 1982 .

[20]  C. J. Hedeskov,et al.  Respiration and glycolysis of normal human lymphocytes. , 1966, Blood.

[21]  C. Munro,et al.  The K veim response: still useful, still a puzzle. , 1987, Thorax.