A novel approach for the selection of human sperm using annexin V-binding and flow cytometry

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dc.contributor.authorHoogendijk C.F.
dc.contributor.authorKruger T.F.
dc.contributor.authorBouic P.J.D.
dc.contributor.authorHenkel R.R.
dc.date.accessioned2011-05-15T16:16:25Z
dc.date.available2011-05-15T16:16:25Z
dc.date.issued2009
dc.description.abstractObjective: To develop a method whereby sperm with phosphatidylserine externalization can be separated from those without this feature. Because annexin V binds phosphatidylserine, this study is using this feature to select functional spermatozoa. In addition, the relationship between annexin V binding in human spermatozoa and normal sperm morphology according to strict criteria was to be assessed. Design: Prospective study. Setting: Department of Obstetrics and Gynaecology of Stellenbosch University at Tygerberg Academic Hospital, Tygerberg, South Africa. Patient(s): Semen from 14 healthy sperm donors. Exclusion criterion was the presence of less than 20 × 106/mL total motile spermatozoa in the original sample. Main Outcome Measure(s): Annexin V-negative sperm, annexin V-positive sperm, normal sperm morphology. Intervention(s): An aliquot of a semen sample after double density gradient centrifugation was incubated with annexin V fluorescein isothiocyanate conjugate (FITC). Cell fluorescence signals were determined using a FACScalibur flow cytometer equipped with a FACSSort fluidic sorting module. The sorting procedure delivered two sperm subpopulations: annexin V-negative and annexin V-positive. Morphology slides were made and the sperm morphology was assessed according to strict criteria. Result(s): There was a significant enrichment of annexin V-negative sperm as well as morphologically normal sperm in the annexin V-negative subgroup after separation with flow cytometry. The percentage of morphologically normal sperm increased from 8.3% in the control to 11.9% in the annexin V-negative fraction, whereas the percentage of annexin V-positive sperm decreased to 5.7%. Conclusion(s): The annexin V-negative sperm subpopulation had morphologically superior quality sperm compared to annexin V-positive sperm. It is important to select morphologically normal sperm during intracytoplasmic sperm injection (ICSI) as it may contribute to increased implantation and pregnancy rates (PR). © 2009 American Society for Reproductive Medicine.
dc.description.versionArticle
dc.identifier.citationFertility and Sterility
dc.identifier.citation91
dc.identifier.citation4
dc.identifier.issn00150282
dc.identifier.other10.1016/j.fertnstert.2008.01.042
dc.identifier.urihttp://hdl.handle.net/10019.1/13773
dc.subjectlipocortin 5
dc.subjectphosphatidylserine
dc.subjectadult
dc.subjectapoptosis
dc.subjectarticle
dc.subjectcontrolled study
dc.subjectflow cytometry
dc.subjecthuman
dc.subjecthuman experiment
dc.subjectmale
dc.subjectnormal human
dc.subjectoutcome assessment
dc.subjectpriority journal
dc.subjectprotein binding
dc.subjectsperm
dc.subjectspermatozoon
dc.subjectAdult
dc.subjectAnnexin A5
dc.subjectCase-Control Studies
dc.subjectCell Separation
dc.subjectFlow Cytometry
dc.subjectHumans
dc.subjectMale
dc.subjectMiddle Aged
dc.subjectProtein Binding
dc.subjectSemen Analysis
dc.subjectSpermatozoa
dc.titleA novel approach for the selection of human sperm using annexin V-binding and flow cytometry
dc.typeArticle
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