2034 POLYMERIZED ALGINIC ACID BEAD AND SPERM INJECTION THROUGH MICRO-MANIPULATION AS A VEHICLE FOR CRYOPRESERVATION AND RECOVERY OF SMALL NUMBERS OF HUMAN SPERM

Abstract

You have accessJournal of UrologyInfertility: Basic Research, Physiology, Pathophysiology1 Apr 20132034 POLYMERIZED ALGINIC ACID BEAD AND SPERM INJECTION THROUGH MICRO-MANIPULATION AS A VEHICLE FOR CRYOPRESERVATION AND RECOVERY OF SMALL NUMBERS OF HUMAN SPERM James Hotaling, Ji Wen, Jessica Belmonte, Craig Niederberger, and Gail S. Prins James HotalingJames Hotaling Chicago, IL More articles by this author , Ji WenJi Wen Shandong, China, People's Republic of More articles by this author , Jessica BelmonteJessica Belmonte Chicago, IL More articles by this author , Craig NiederbergerCraig Niederberger Chicago, IL More articles by this author , and Gail S. PrinsGail S. Prins Chicago, IL More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2013.02.2453AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES While routine cryopreservation of human sperm is effective for normozoospermic specimens, severe oligozoospermic samples are limited by the marked reductions in sperm motility and viability post-thaw. This is exacerbated in TESE when retrieval can yield <1000 sperm. Alternative methods are required that enable sufficient post-thaw sperm recovery, ideally in several aliquots, to permit multiple ICSI attempts. Herein, we describe a novel technique involving microinjection of limited sperm numbers into pre-formed polymerized alginate beads that can be frozen and thawed individually. METHODS Semen samples were collected from normospermic men (n=10), sperm were washed in Ham's F10 and a portion was frozen using standard cryoprotocol. Fresh alginate beads were formed by dropping 10 ul 7.3 mg/ml alginic acid into 135 mM CaCl2. Spermatozoa (n=10 to 1000) were aspirated and injected into individual alginate beads using a Nikon-Narigishe micromanipulator. Individual beads were transferred to freezing medium in 0.5 ml vials and frozen in liquid nitrogen. Following thaw, beads were solubilized in Na-citrate and sperm recovered by micromanipulator aspiration to fresh Ham'sF-10. Post-thaw sperm recovery, viability and motility were assessed for both cryopreservation methods and compared to fresh parameters. RESULTS Greater than 90% of injected spermatozoa were recovered from alginate beads post-thaw supporting the efficacy of this technique for successful cryopreservation of limited sperm. There was a significant post-thaw decline in sperm motility and viability using alginate bead or standard cryopreservation as compared to neat semen, respectively (Motility: 21.97%, 36.45% vs 69.40%; Viability: 48.78%, 51.33% vs 80.45%). The significant decrease in motility (21.97% vs 36.45% motile post-thaw, P<0.05) between alginate cryopreservation vs standard protocol was counter-balanced by higher viability of the immotile sperm in alginate beads (26.81% vs 14.93%, P<0.05). There was no difference in sperm viability post-thaw using either cryoprotocol (alginate: 48.78% vs standard: 51.33%, P>0.05). CONCLUSIONS Microinjection of sperm into pre-made alginate beads yields equivalent viable sperm post-thaw as standard cryopreservation. Coupled with high post-thaw yield, this approach provides a previously unrealized opportunity for fertility preservation in men with limited sperm numbers recovered by TESE and may permit multiple ICSI attempts from a single tissue sample obviating further surgery. © 2013 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 189Issue 4SApril 2013Page: e835 Advertisement Copyright & Permissions© 2013 by American Urological Association Education and Research, Inc.MetricsAuthor Information James Hotaling Chicago, IL More articles by this author Ji Wen Shandong, China, People's Republic of More articles by this author Jessica Belmonte Chicago, IL More articles by this author Craig Niederberger Chicago, IL More articles by this author Gail S. Prins Chicago, IL More articles by this author Expand All Advertisement Advertisement PDF DownloadLoading ...