Difference between revisions of "Kalbacova 2003 Cytometry"
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{{Publication | {{Publication | ||
|title= | |title=Kalbácová M, Vrbacký M, Drahota Z, Melková 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:110-6. | ||
|authors=Kalbacova M, Vrbacky M, Drahota Z, Melkova Z | |info=[http://www.ncbi.nlm.nih.gov/pubmed/12655654 PMID: 12655654 Open Access] | ||
|authors=Kalbacova M, Vrbacky M, Drahota Z, Melkova Z | |||
|year=2003 | |year=2003 | ||
|abstract=Background: Determination of mitochondrial membrane | |journal=Cytometry | ||
potential ((m) is widely used to characterize cellular | |abstract='''Background:''' Determination of mitochondrial membrane potential ((m) is widely used to characterize cellular metabolism, viability, and apoptosis. Changes of ΔΨm induced by inhibitors of oxidative phosphorylation characterize | ||
metabolism, viability, and apoptosis. Changes of | respective contributions of mitochondria and glycolysis to adenosine triphosphate (ATP) synthesis. | ||
induced by inhibitors of oxidative phosphorylation characterize | '''Methods:''' ΔΨm 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 ΔΨ-dihexyloxacarbocyanine iodide | ||
respective contributions of mitochondria and glycolysis | (DiOC6(3)), tetramethylrhodamine ethyl ester, and Mito-Tracker Red. Mitochondrial function was further characterized by oxygen consumption. | ||
to adenosine triphosphate (ATP) synthesis. | '''Results:''' Inhibition of respiration by antimycin A or uncoupling | ||
Methods: | of mitochondria by FCCP decreased ΔΨm in both cell lines. Inhibition of ATP production by oligomycin or atractyloside induced a moderate decrease of ΔΨm | ||
determined by flow cytometry and spectrofluorometry. Its | in HeLa G cells and an increase of ΔΨm in BSC-40 cells. Statistically significant differences in ΔΨm between the two cell lines were found with both flow cytometry and spectrofluorometry. Respirometry showed higher basal | ||
changes induced by specific mitochondrial inhibitors | |||
were evaluated using 3, | |||
(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 | |||
both cell lines. Inhibition of ATP production by oligomycin | |||
or atractyloside induced a moderate decrease of | |||
in HeLa G cells and an increase of | |||
Statistically significant differences in | |||
two cell lines were found with both flow cytometry and | |||
spectrofluorometry. Respirometry showed higher basal | |||
and FCCP-stimulated respiration in BSC-40 cells. | and FCCP-stimulated respiration in BSC-40 cells. | ||
Conclusion: Changes of | '''Conclusion:''' Changes of ΔΨm 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 | ||
showed that BSC-40 cells are more sensitive than HeLa G | aerobic ATP production. Determination of ΔΨm changes by flow cytometry exhibited greater sensitivity than the ones by spectrofluorometry. | ||
cells to inhibitors of mitochondrial function, suggesting | |keywords=Green monkey kidney BSC-40, Human cervical carcinoma HeLa G cell lines | ||
that BSC-40 cells are more dependent than HeLa G cells on | |mipnetlab=CZ Prague Houstek J, CZ Hradec Kralove Cervinkova Z | ||
aerobic ATP production. Determination of | |||
by flow cytometry exhibited greater sensitivity than the | |||
ones by spectrofluorometry. | |||
| | |||
}} | }} | ||
{{Labeling | {{Labeling | ||
| | |area=Respiration, Instruments;methods | ||
|tissues= | |tissues=Other cell lines | ||
| | |preparations=Intact cells | ||
| | |couplingstates=ROUTINE | ||
|instruments=Oxygraph-2k | |instruments=Oxygraph-2k | ||
| | |additional=Spectrophotometry; Spectrofluorimetry, monkey | ||
}} | }} | ||
Latest revision as of 11:57, 9 November 2016
| Kalbácová M, Vrbacký M, Drahota Z, Melková 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:110-6. |
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 ΔΨm induced by inhibitors of oxidative phosphorylation characterize respective contributions of mitochondria and glycolysis to adenosine triphosphate (ATP) synthesis. Methods: ΔΨm 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 ΔΨ-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 ΔΨm in both cell lines. Inhibition of ATP production by oligomycin or atractyloside induced a moderate decrease of ΔΨm in HeLa G cells and an increase of ΔΨm in BSC-40 cells. Statistically significant differences in ΔΨm 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 ΔΨm 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 ΔΨm changes by flow cytometry exhibited greater sensitivity than the ones by spectrofluorometry. • Keywords: Green monkey kidney BSC-40, Human cervical carcinoma HeLa G cell lines
• O2k-Network Lab: CZ Prague Houstek J, CZ Hradec Kralove Cervinkova Z
Labels: MiParea: Respiration, Instruments;methods
Tissue;cell: Other cell lines Preparation: Intact cells
Coupling state: ROUTINE
HRR: Oxygraph-2k
Spectrophotometry; Spectrofluorimetry, monkey
