2045 SUSPENDED ANIMATION OF HUMAN SPERM: ULTRA-RAPID VITRIFICATION USING A NOVEL MICROCAPILLARY SYSTEM

Abstract

You have accessJournal of UrologyInfertility: Basic Research, Physiology, Pathophysiology1 Apr 20132045 SUSPENDED ANIMATION OF HUMAN SPERM: ULTRA-RAPID VITRIFICATION USING A NOVEL MICROCAPILLARY SYSTEM Yahir Santiago-Lastra, Joseph W. McQuaid, Diane Wright, Mehmet Toner, Cigdem Tanrikut, and Thomas L. Toth Yahir Santiago-LastraYahir Santiago-Lastra Boston, MA More articles by this author , Joseph W. McQuaidJoseph W. McQuaid Boston, MA More articles by this author , Diane WrightDiane Wright Boston, MA More articles by this author , Mehmet TonerMehmet Toner Boston, MA More articles by this author , Cigdem TanrikutCigdem Tanrikut Boston, MA More articles by this author , and Thomas L. TothThomas L. Toth Boston, MA More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2013.02.2464AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Freezing very low concentrations of sperm remains a challenge. Conventional slow-freeze methods require dilution with relatively large volumes and high concentrations of traditional cryoprotectants with potentially lethal osmotic and cytotoxic effects. Development of ultra-rapid vitrification techniques allows for freezing of very low volumes using low concentrations of nontoxic cryoprotectants, such as sugars, in addition to very low doses of permeable cryoprotectant. Our investigation was undertaken to determine the feasibility of ultra-rapid freezing via vitrification of human sperm samples with 0.5M trehalose and 3.0M propanediol (PROH) using a novel microcapillary system. METHODS 84 fresh ejaculated samples were obtained from healthy men and prepared using the swim-up method in human tubal fluid (HTF) media supplemented with 5% human serum albumin (HSA). All samples had at least 20 million spermatozoa/ml and at least 50% motility prior to swim-up preparation. These samples were then diluted to varying concentrations and divided into equal volumes for vitrification into the following groups: (1) 0.25M trehalose (a natural sugar) to act as an impermeable cryoprotectant, (2) vitrification with 0.25M trehalose and propanediol (PROH), (3) vitrification without cryoprotectant, or (4) no freezing/vitrification (control). Each sample was loaded into a 100μ microcapillary and the ends were heat-sealed to create a closed system. The samples undergoing vitrification were plunged into a liquid nitrogen bath for 20 seconds and then warmed in a 37°C solution of phosphate-buffered saline. Each sample was expelled from the capillaries and motility was qualitatively assessed via microscopic visualization. All samples were further assessed by computer-assisted semen analysis. RESULTS Preliminary results demonstrate consistent and statistically significant (p<0.05) recovery of motile sperm in samples that were vitrified in 100μ microcapillaries with HTF media plus 5% HSA supplemented by 0.25M trehalose and PROH. CONCLUSIONS Ultra-rapid vitrification of human sperm samples in microcapillaries is feasible using low volumes of a nontoxic sugar, such as trehalose, as an alternative to traditional toxic cryoprotectants. This is a particularly relevant issue with men who produce very little sperm in the ejaculate and in men who undergo any variety of surgical sperm extraction efforts for either obstructive or nonobstructive azoospermia. © 2013 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 189Issue 4SApril 2013Page: e839 Advertisement Copyright & Permissions© 2013 by American Urological Association Education and Research, Inc.MetricsAuthor Information Yahir Santiago-Lastra Boston, MA More articles by this author Joseph W. McQuaid Boston, MA More articles by this author Diane Wright Boston, MA More articles by this author Mehmet Toner Boston, MA More articles by this author Cigdem Tanrikut Boston, MA More articles by this author Thomas L. Toth Boston, MA More articles by this author Expand All Advertisement Advertisement PDF DownloadLoading ...