Kunz-Schughart, L. A. and Habbersett, RC and Freyer, JP (1997) Mitochondrial function in oncogene-transfected rat fibroblasts isolated from multicellular spheroids. AMERICAN JOURNAL OF PHYSIOLOGY-CELL PHYSIOLOGY, 273 (5). C1487-C1495. ISSN 0363-6143, 1522-1563
Full text not available from this repository.Abstract
Two mitochondrion-specific fluorochromes, 10-N-nonyl acridine orange (NAO) and rhodamine 123 (Rh123), were used to determine the mechanism responsible for alterations in energy metabolism of transformed rat embryo fibroblast cells isolated from different locations within multicellular spheroids. Accumulation of Rh123 depends on intact mitochondrial membrane potential, whereas NAO is taken up by mitochondria independently of their function and thus represents mitochondrial distribution only. A reproducible selective dissociation procedure was used to isolate cells from different locations within the spheroids. After isolation, cells were simultaneously stained with one mitochondrial stain and the DNA dye Hoechst 33342, and several parameters, including cell volume, were monitored via multilaser-multiparameter flow cytometry. Our data clearly show a decrease in the uptake of Rh123 in cells from the periphery to the inner regions of the tumor spheroids, reflecting a persistent alteration in mitochondrial function. However, NAO staining experiments showed no reduction in the total mitochondrial mass per unit cell volume. Because cells were exposed to stain under uniform conditions after isolation from the spheroid, these data indicate that downregulation of mitochondrial function is associated with cell quiescence rather than a transient effect of reduced nutrient availability. This result, which is in accordance with data from two other cell lines (EMT6 and 9L), might reflect a general phenomenon in multicellular spheroids, supporting the hypothesis that quiescent cells in the innermost viable spheroid layer stably reduce their mitochondrial function, presumably to compensate for lower nutrient supply and/or decreased energy demand.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | NUCLEAR-MAGNETIC-RESONANCE; OXYGEN-CONSUMPTION; DIFFERENT REGIONS; TUMOR SPHEROIDS; SELECTIVE DISSOCIATION; METABOLIC ALTERATIONS; ACRIDINE-ORANGE; CELLS; GROWTH; RHODAMINE-123; rat embryo fibroblasts; mitochondria; cell cycle; flow cytometry |
| Subjects: | 500 Science > 570 Life sciences 600 Technology > 610 Medical sciences Medicine |
| Divisions: | Medicine > Lehrstuhl für Pathologie |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 02 Mar 2023 09:05 |
| Last Modified: | 02 Mar 2023 09:05 |
| URI: | https://pred.uni-regensburg.de/id/eprint/50417 |
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