Study of mitochondrial membrane potential, reactive oxygen species, DNA fragmentation and cell viability by flow cytometry in human sperm.
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 (Delta Psi(m)), generation of reactive oxygen species, DNA fragmentation and cell viability.


RESULTS
In neat semen, a positive correlation was found between the percentage of Delta Psi(m)(high) sperm cells and standard semen parameters (concentration/motility). Sperm cells depicting Delta Psi(m)(high) 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 Delta Psi(m), with Delta Psi(m)(high) sperm positively correlated with forward motility and also with high fertilization rates after IVF.


CONCLUSIONS
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.
Marchetti
Carole
C
INSERM U459, Faculté de Médecine 1, Hôpital Jeanne de Flandre, Lille Cedex, France.
Obert
Guillaume
G
Deffosez
André
A
Formstecher
Pierre
P
Marchetti
Philippe
P
D000328:Adult; D002470:Cell Survival / Q000502:physiology; D053938:DNA Fragmentation; D005306:Fertilization; D005307:Fertilization in Vitro; D005434:Flow Cytometry; D006801:Humans; D007248:Infertility, Male / Q000503:physiopathology*; D008297:Male; D008564:Membrane Potentials / Q000502:physiology; D008928:Mitochondria / Q000502:physiology*; D017382:Reactive Oxygen Species / Q000378:metabolism*; D013076:Sperm Count; D013081:Sperm Motility; D013094:Spermatozoa / Q000502:physiology*
D017382:Reactive Oxygen Species