Kalbacova 2003 Cytometry
Kalbacova M, Vrbacky M, Drahota Z, Melkova Z (2003) Comparison of the effect of mitochondrial inhibitors on mitochondrial membrane potential in two different cell lines using flow cytometry and spectrofluorometry. Cytometry 52A: 110-116. |
Kalbacova M, Vrbacky M, Drahota Z, Melkova Z (2003) Cytometry
Abstract: Background: Determination of mitochondrial membrane potential ((m) is widely used to characterize cellular metabolism, viability, and apoptosis. Changes of ΞΞ¨m induced by inhibitors of oxidative phosphorylation characterize respective contributions of mitochondria and glycolysis to adenosine triphosphate (ATP) synthesis. Methods: ΞΞ¨m in BSC-40 and HeLa G cell lines was determined by flow cytometry and spectrofluorometry. Its changes induced by specific mitochondrial inhibitors were evaluated using 3,3 ΞΞ¨-dihexyloxacarbocyanine iodide (DiOC6(3)), tetramethylrhodamine ethyl ester, and Mito-Tracker Red. Mitochondrial function was further characterized by oxygen consumption. Results: Inhibition of respiration by antimycin A or uncoupling of mitochondria by FCCP decreased ΞΞ¨m in both cell lines. Inhibition of ATP production by oligomycin or atractyloside induced a moderate decrease of ΞΞ¨m in HeLa G cells and an increase of ΞΞ¨m in BSC-40 cells. Statistically significant differences in ΞΞ¨m between the two cell lines were found with both flow cytometry and spectrofluorometry. Respirometry showed higher basal and FCCP-stimulated respiration in BSC-40 cells. Conclusion: Changes of ΞΞ¨m and oxygen consumption showed that BSC-40 cells are more sensitive than HeLa G cells to inhibitors of mitochondrial function, suggesting that BSC-40 cells are more dependent than HeLa G cells on aerobic ATP production. Determination of ΞΞ¨m changes by flow cytometry exhibited greater sensitivity than the ones by spectrofluorometry. β’ Keywords: green monkey kidney BSC-40, human cervical carcinoma HeLa G cell lines
β’ O2k-Network Lab: CZ_Prague_Houstek J
Labels:
Stress:Cancer; Apoptosis; Cytochrome c"Cancer; Apoptosis; Cytochrome c" is not in the list (Cell death, Cryopreservation, Ischemia-reperfusion, Permeability transition, Oxidative stress;RONS, Temperature, Hypoxia, Mitochondrial disease) of allowed values for the "Stress" property. Organism: Human, Other Mammal"Other Mammal" is not in the list (Human, Pig, Mouse, Rat, Guinea pig, Bovines, Horse, Dog, Rabbit, Cat, ...) of allowed values for the "Mammal and model" property. Tissue;cell: Kidney Preparation: Intact Cell; Cultured; Primary"Intact Cell; Cultured; Primary" is not in the list (Intact organism, Intact organ, Permeabilized cells, Permeabilized tissue, Homogenate, Isolated mitochondria, SMP, Chloroplasts, Enzyme, Oxidase;biochemical oxidation, ...) of allowed values for the "Preparation" property.
Regulation: Coupling; Membrane Potential"Coupling; Membrane Potential" is not in the list (Aerobic glycolysis, ADP, ATP, ATP production, AMP, Calcium, Coupling efficiency;uncoupling, Cyt c, Flux control, Inhibitor, ...) of allowed values for the "Respiration and regulation" property.
HRR: Oxygraph-2k
Spectrophotometry; Spectrofluorimetry, monkey