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Andrology Lab Corner^*

Utility of Magnetic Cell Separation as a Molecular Sperm Preparation Technique

ASHOK AGARWAL

MACIEJ ZBOROWSKI

SONJA GRUNEWALD

HANS-JUERGEN GLANDER

UWE PAASCH

From the ^* Department of Andrology and Reproductive Tissue Banking, The Toronto Institute for Reproductive Medicine (ReproMed), Toronto, Canada; the Center for Advanced Research in Human Reproduction, Infertility, and Sexual Function, Glickman Urological and Kidney Institute and the Department of Biomedical Engineering, Lerner Research Institute, The Cleveland Clinic, Cleveland, Ohio; and the Department of Dermatology and Andrology, University of Leipzig, Leipzig, Germany.

Correspondence to: Prof Dr Uwe Paasch, Department of Dermatology, Venereology, and Allergology, European Training Center of the European Academy of Andrology, University of Leipzig, 04103 Leipzig, Germany (e-mail: uwe.paasch{at}medizin.uni-leipzig.de).

Abstract

Assisted reproductive techniques (ARTs) have become the treatment of choice in many cases of infertility; however, the current success rates of these procedures remain suboptimal. Programmed cell death (apoptosis) most likely contributes to failed ART and to the decrease in sperm quality after cryopreservation. There is a likelihood that some sperm selected for ART will display features of apoptosis despite their normal appearance, which may be partially responsible for the low fertilization and implantation rates seen with ART. One of the features of apoptosis is the externalization of phosphatidylserine (PS) residues, which are normally present on the inner leaflet of the sperm plasma membrane. Colloidal superparamagnetic microbeads (50 nm in diameter) conjugated with annexin V bind to PS and are used to separate dead and apoptotic spermatozoa by magnetic-activated cell sorting (MACS). Cells with externalized PS will bind to these microbeads, whereas nonapoptotic cells with intact membranes do not bind and could be used during ARTs. We have conducted a series of experiments to investigate whether the MACS technology could be used to improve ART outcomes. Our results clearly indicate that integrating MACS as a part of sperm preparation techniques will improve semen quality and cryosurvival rates by eliminating apoptotic sperm. Nonapoptotic spermatozoa prepared by MACS display higher quality in terms of routine sperm parameters and apoptosis markers. The higher sperm quality is represented by an increased oocyte penetration potential and cryosurvival rates. Thus, the selection of nonapoptotic spermatozoa by MACS should be considered to enhance ART success rates.

     Key words: Annexin V, apoptosis, cryopreservation, human, MACS, male infertility

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