Kalbacova 2003 Cytometry: Difference between revisions
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'''Conclusion:''' Changes of ฮฮจm 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 | '''Conclusion:''' Changes of ฮฮจm 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 ฮฮจm changes by flow cytometry exhibited greater sensitivity than the ones by spectrofluorometry. | aerobic ATP production. Determination of ฮฮจm changes by flow cytometry exhibited greater sensitivity than the ones by spectrofluorometry. | ||
|keywords= | |keywords=Green monkey kidney BSC-40, Human cervical carcinoma HeLa G cell lines | ||
|mipnetlab=CZ_Prague_Houstek J, CZ Prague Bioenergetics | |mipnetlab=CZ_Prague_Houstek J, CZ Prague Bioenergetics | ||
}} | }} | ||
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|instruments=Oxygraph-2k | |instruments=Oxygraph-2k | ||
|injuries=Cancer; Apoptosis; Cytochrome c | |injuries=Cancer; Apoptosis; Cytochrome c | ||
| | |model cell lines=Other cell lines | ||
|preparations=Intact Cell; Cultured; Primary | |preparations=Intact Cell; Cultured; Primary | ||
|couplingstates=ROUTINE | |||
|additional=Spectrophotometry; Spectrofluorimetry, monkey | |additional=Spectrophotometry; Spectrofluorimetry, monkey | ||
}} | }} | ||
Revision as of 17:41, 12 February 2013
| 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 ฮฮจm induced by inhibitors of oxidative phosphorylation characterize respective contributions of mitochondria and glycolysis to adenosine triphosphate (ATP) synthesis. Methods: ฮฮจm 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 ฮฮจ-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 ฮฮจm in both cell lines. Inhibition of ATP production by oligomycin or atractyloside induced a moderate decrease of ฮฮจm in HeLa G cells and an increase of ฮฮจm in BSC-40 cells. Statistically significant differences in ฮฮจm 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 ฮฮจm 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 ฮฮจm changes by flow cytometry exhibited greater sensitivity than the ones by spectrofluorometry. โข Keywords: Green monkey kidney BSC-40, Human cervical carcinoma HeLa G cell lines
โข O2k-Network Lab: CZ_Prague_Houstek J, CZ Prague Bioenergetics
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.
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.
Coupling state: ROUTINE
HRR: Oxygraph-2k
Spectrophotometry; Spectrofluorimetry, monkey
