HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) All Versions of this Article: 74/3/530    most recent biolreprod.105.046607v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal My Folders Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Said, T. Articles by Paasch, U. Search for Related Content PubMed PubMed Citation Articles by Said, T. Articles by Paasch, U. Agricola Articles by Said, T. Articles by Paasch, U.

Selection of Nonapoptotic Spermatozoa As a New Tool for Enhancing Assisted Reproduction Outcomes: An In Vitro Model¹

Center for Advanced Research in Human Reproduction, Infertility and Sexual Function;³ Glickman Urological Institute;⁴ Department of Obstetrics-Gynecology;⁵ Department of Biostatistics and Epidemiology,⁶ Cleveland Clinic Foundation, Cleveland, Ohio 44195 Department of Dermatology/Andrology Unit,⁷ University of Leipzig, 04103 Leipzig, Germany

ABSTRACT

Magnetic cell sorting (MACS) using annexin V-conjugated microbeads eliminates apoptotic spermatozoa based on the externalization of phosphatidylserine residues. The procedure delivers two sperm fractions: annexin V-negative (nonapoptotic) and annexin V-positive (apoptotic). Our aim was to determine whether the sperm fertilizing potential can be improved by selecting a nonapoptotic fraction using MACS. Semen samples (n = 35) were subjected to separation on a density gradient followed by MACS. Extent of apoptosis was assessed by measuring levels of activated caspase 3 using fluorescein-labeled inhibitors of caspase, alterations in mitochondrial membrane potential (MMP) using a lipophilic cationic dye, and DNA fragmentation using terminal deoxynucleotidyl transferase-mediated fluorescein-dUTP nick end labeling assay. The sperm fertilization potential was assessed using hamster oocyte penetration assay and hamster oocyte-intracytoplasmic sperm injection (ICSI). Annexin V-negative sperm displayed superior quality in terms of high motility, low caspase 3 activation, MMP integrity, and small extent of DNA fragmentation. Annexin V-negative sperm demonstrated higher oocyte penetration capacity but comparable sperm chromatin decondensation (SCD) following ICSI. Conversely, the annexin V-positive sperm presented with poor quality and fertilization potential. The oocyte penetration rate was negatively correlated with apoptotic marker expression, whereas SCD following ICSI was only associated with apoptosis on sperm-damaged membranes. We conclude that apoptosis appears to impact sperm-oocyte penetration rate; however, it does not seem to affect early stages of fertilization such as SCD in spermatozoa of healthy donors. The selection of nonapoptotic sperm by MACS may be used to enhance results of in vitro fertilization by increasing sperm-oocyte penetration.

apoptosis, in vitro fertilization, sperm

² Correspondence: Uwe Paasch, University of Leipzig, Department of Dermatology, Training Center of the European Academy of Andrology, Stephanstrasse 11, 04103 Leipzig, Germany. FAX: 49 341 9718649; uwe.paasch{at}medizin.uni-leipzig.de

¹ Supported by funds from the Cleveland Clinic Foundation (RPC7812), the German Research Council (Deutsche Forschungsgemeinschaft, DFG GL 199/4), and a Junior Research Grant from the Faculty of Medicine, University of Leipzig (formel.1–54).

This article has been cited by other articles:

				T. M. Said, A. Agarwal, M. Zborowski, S. Grunewald, H.-J. Glander, and U. Paasch Utility of Magnetic Cell Separation as a Molecular Sperm Preparation Technique J Androl, March 1, 2008; 29(2): 134 - 142. [Abstract] [Full Text] [PDF]

				M. Bungum, M. Spano, P. Humaidan, P. Eleuteri, M. Rescia, and A. Giwercman Sperm chromatin structure assay parameters measured after density gradient centrifugation are not predictive for the outcome of ART Hum. Reprod., January 1, 2008; 23(1): 4 - 10. [Abstract] [Full Text] [PDF]

				J. Adjaye, R. Herwig, T. C. Brink, D. Herrmann, B. Greber, S. Sudheer, D. Groth, J. W. Carnwath, H. Lehrach, and H. Niemann Conserved molecular portraits of bovine and human blastocysts as a consequence of the transition from maternal to embryonic control of gene expression Physiol Genomics, October 19, 2007; 31(2): 315 - 327. [Abstract] [Full Text] [PDF]

				E. Marti, R. Perez-Pe, T. Muino-Blanco, and J. A. Cebrian-Perez Comparative Study of Four Different Sperm Washing Methods Using Apoptotic Markers in Ram Spermatozoa J Androl, November 1, 2006; 27(6): 746 - 753. [Abstract] [Full Text] [PDF]