Study of mitochondrial membrane potential, reactive oxygen species, DNA fragmentation and cell viability by flow cytometry in human sperm

• Published in: Human reproduction (Oxford) Vol. 17; no. 5; pp. 1257 - 1265
• Main Authors: Marchetti, Carole, Obert, Guillaume, Deffosez, André, Formstecher, Pierre, Marchetti, Philippe
• Format: Journal Article
• Language: English
• Published: Oxford Oxford University Press 01-05-2002
• Subjects: Adult; Biological and medical sciences; cell death; Cell Survival - physiology; DNA Fragmentation; Fertilization; Fertilization in Vitro; Flow Cytometry; Fundamental and applied biological sciences. Psychology; Humans; infertility; Infertility, Male - physiopathology; Male; Mammalian reproduction. General aspects; Membrane Potentials - physiology; mitochondria; Mitochondria - physiology; Reactive Oxygen Species - metabolism; sperm; Sperm Count; Sperm Motility; Spermatozoa - physiology; Vertebrates: reproduction; sperm; cell death; infertility; mitochondria; DNA fragmentation; Human; Flow cytometry; Spermatozoa; Motility; Prediction; Electrophysiology; Germinal cell; Free radical; Mitochondria; Cell death; DNA; Male sterility; Membrane potential; Male genital diseases
• ISSN: 0268-1161; 1460-2350; 1460-2350
• DOI: 10.1093/humrep/17.5.1257

BACKGROUND: Sperm cell death appears to be a cause of male infertility. The objective of this study was to determine the most reliable method for the evaluation of sperm quality in semen samples during sperm preparation for IVF. METHODS: Conventional analysis of semen samples was compared with several cytofluorometric methods detecting death-associated changes. Neat semen from infertile patients and sperm prepared by PureSperm® gradient were studied by conventional microscopy and analysed for mitochondrial membrane potential (ΔΨΨm), generation of reactive oxygen species, DNA fragmentation and cell viability. RESULTS: In neat semen, a positive correlation was found between the percentage of ΔΨΨmhigh sperm cells and standard semen parameters (concentration/motility). Sperm cells depicting ΔΨΨmhigh and cells with low DNA fragmentation displayed high fertilization rate after IVF. The only changes that could be detected in prepared sperm were changes in ΔΨΨm, with ΔΨΨmhigh sperm positively correlated with forward motility and also with high fertilization rates after IVF. CONCLUSION: Analysis of mitochondrial membrane potential is the most sensitive test by which to determine sperm quality. These findings promise development of a test that may help to predict successful IVF.