0000000000448644
AUTHOR
Ulrich Karbach
Proliferation-associated oxygen consumption and morphology of tumor cells in monolayer and spheroid culture.
The oxygen consumption rate, proliferative activity, and morphology of EMT6/Ro mouse mammary sarcoma cells in monolayer and multicellular spheroid culture have been investigated in a comparative study. During the transition of monolayer cells from the exponential into the plateau growth phase, there is a distinct decrease in the cellular volume that is associated with a corresponding decrease in the proliferative and respiratory activity of the cells. The decline in cell volume is mainly due to a decrease in the content of cytoplasm, whereas the size of the nucleus is only slightly reduced. A concomitant decrease in the number of mitochondria per cell obviously accounts for the reduction in…
Interrelationship Among Morphology, Metabolism, and Proliferation of Tumor Cells in Monolayer and Spheroid Culture
Previous investigations have indicated a positive correlation between the proliferative and metabolic activities of tumor cells in monolayer culture (Freyer et al., 1984; Freyer and Sutherland, 1985; Walenta and Mueller-Klieser, 1987). On the other hand, no difference in the local oxygen consumption has been found between highly proliferating outer cell areas and non-proliferating inner cellular regions in multicellular tumor spheroids (Mueller-Klieser, 1984, 1987). Therefore, the interrelationship among metabolism, proliferation, and cellular morphology was investigated systematically in tumor cells both in monolayer and spheroid culture.
Pathophysiological approaches to identifying tumor hypoxia in patients
The present report summarizes observations of the authors on tumor oxygenation and on techniques for characterizing tumor hypoxia in patients. Cryospectrophotometric measurements of HbO2 saturations in tumor microvessels allow for estimates of the proportion of well oxygenated tissue regions. Labeling of tissue areas at oxygen (O2) tensions (pO2) less than 10 mm Hg with misonidazole may be used for a general characterization of the oxygenation status in patient tumors rather than for the determination of the radiobiologically hypoxic cell fraction. Quantitative bioluminescence and single photon imaging make it possible to determine ATP concentrations in absolute terms with a spatial resolut…
Calibration of misonidazole labeling by simultaneous measurement of oxygen tension and labeling density in multicellular spheroids
To correlate misonidazole concentrations and oxygen pressures (Po2) at identical locations within EMT6/Ro multi-cell spheroids (mean diameters +/- SD: 867 +/- 20 microns), Po2 measurements were performed with oxygen-sensitive microelectrodes during incubation of these spheroids with tritiated misonidazole (10 mg/I; 445 microCi/mg). In each individual spheroid, Po2 profiles were correlated with the corresponding spatial distribution of misonidazole as quantified by conventional autoradiography and grain counting. To compare the oxygenation status of spheroids in the measuring chamber with that of spheroids in spinner culture, misonidazole labeling was performed in both environments following…
Oxygenation Status of Rhabdomyosarcoma Spheroids with Different Stages of Differentiation
Multicellular tumor spheroids with regional heterogeneities of proliferation and substrate concentrations have been investigated recently to study the biological properties of small tumor nodules prior to onset of vascularization (for reviews see: Mueller-Klieser, 1987; Sutherland, 1988).