Human Reproduction, Vol. 17, No. 5, 1257-1265,
May 2002
© 2002 European Society of Human Reproduction and Embryology
Study of mitochondrial membrane potential, reactive oxygen species, DNA fragmentation and cell viability by flow cytometry in human sperm
1 INSERM U459, Faculté de Médecine 1 and 2 Laboratoire Histologie-Biologie de la Reproduction, Hôpital Jeanne de Flandre, Lille Cedex, France
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 (
m), generation of reactive oxygen species, DNA fragmentation and cell viability. RESULTS: In neat semen, a positive correlation was found between the percentage of mhigh sperm cells and standard semen parameters (concentration/motility). Sperm cells depicting mhigh 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 m, with mhigh 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.
Key words: cell death/DNA fragmentation/infertility/mitochondria/sperm
3 To whom correspondence should be addressed at: INSERM U 459, Faculté de Médecine 1, Place Verdun, F- 59045 Lille Cedex, France. E-mail: philippe.marchetti{at}lille.inserm.fr
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