Kalbacova 2003 Cytometry
|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.|
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
Labels: MiParea: Respiration, Instruments;methods
Tissue;cell: Other cell lines Preparation: Intact cells
Coupling state: ROUTINE
Spectrophotometry; Spectrofluorimetry, monkey