MP66-15 ADVANCES IN THE ULTRA-RAPID VITRIFICATION OF HUMAN SPERMATAZOA WITH A MICROCAPILLARY SYSTEM

Abstract

You have accessJournal of UrologyInfertility: Basic Research, Physiology & Pathophysiology1 Apr 2014MP66-15 ADVANCES IN THE ULTRA-RAPID VITRIFICATION OF HUMAN SPERMATAZOA WITH A MICROCAPILLARY SYSTEM Joseph W. McQuaid, Yahir Santiago-Lastra, Diane Wright, Mehmet Toner, Thomas L. Toth, and Cigdem Tanrikut Joseph W. McQuaidJoseph W. McQuaid More articles by this author , Yahir Santiago-LastraYahir Santiago-Lastra More articles by this author , Diane WrightDiane Wright More articles by this author , Mehmet TonerMehmet Toner More articles by this author , Thomas L. TothThomas L. Toth More articles by this author , and Cigdem TanrikutCigdem Tanrikut More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2014.02.2062AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Freezing low concentrations of sperm from severely oligospermic patients while maintaining adequate post-thaw motility remains a challenge. We previously reported the feasibility of ultra-rapid vitrification using a microcapillary system. In an attempt to improve these results, several modifications were introduced including a larger capillary system and lower concentrations of non-toxic cryoprotectants: trehalose and propanediol. METHODS Using an IRB-approved protocol, 27 fresh ejaculate samples from healthy men were obtained and prepared via a swim-up protocol in human tubal fluid (HTF). 15 samples with an average concentration of 77 million/mL and pre-thaw total motility of 86% were included in our modified protocol. These samples were diluted in a drop-wise, 1:1 fashion with HTF media containing trehalose (0.05M) and propanediol (PrOH, 0.75M). After loading the samples into a series of 200 μm capillaries with heat-sealed ends, they were divided into two equal groups: 1) Control, from which the samples were immediately expelled, washed, then assessed via computer-assisted semen analysis, and 2) Experimental, which was vitrified in a super-cooled, liquid nitrogen bath, stored for up to 24 hours, then thawed, washed, and analyzed. A subset of specimens from the experimental group was used for viability staining. RESULTS Consistent post-thaw motility was achieved using this protocol. While a total motility of 48.6% was recorded in the control, the experimental group demonstrated an overall post-thaw motility of 10.5%. Subset analysis allowed stratification of samples based upon their ability to withstand this process. Of the highest-performing specimens, defined as those with a post-thaw motility >15%, we were able to conserve 22% of the sham control motility in our vitrified samples. CONCLUSIONS Building upon our prior experience, we sought to enhance sperm survival and improve post-thaw motility. Modifications included use of a larger capillary to reduce shear stress, conversion to a less viscous and less toxic media, and incorporation of a wash prior to final analysis. Gains were achieved in post-thaw motility and confirmed the feasibility of this system. However, it is clear from viability stains and subset analysis that there are yet undiscovered intrinsic and extrinsic variables affecting optimal recovery of specimens with this process. We will continue efforts to optimize this system for use in severely oligoospermic patients. © 2014FiguresReferencesRelatedDetails Volume 191Issue 4SApril 2014Page: e746 Advertisement Copyright & Permissions© 2014MetricsAuthor Information Joseph W. McQuaid More articles by this author Yahir Santiago-Lastra More articles by this author Diane Wright More articles by this author Mehmet Toner More articles by this author Thomas L. Toth More articles by this author Cigdem Tanrikut More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...