Abstract
The aim of this study was to determine if the addition of HA to a BL vitrification (VTF) solution promotes membrane stabilization of cryopreserved cellular tissue. Prospective, randomized study. In 2011, we began prospectively vitrified PGS cases with (+) or without (-) HA for Day 6 or D5 BLs, respectively. In 2012, we randomly assigned all PGS cases, by patient, to VTF +HA or -HA. Aseptic microSecure (μS)–VTF was applied to all PGS-BLs using ICE VTF-BL solutions/3-step dilution. μS-VTF is a FDA compliant method that offers superior QC characteristics, including high-security weld seals, in-situ vapor storage, and dual-colored, tamperproof labeling. Thirty warming cycles were conducted and chi-squared analysis was performed to compare survival rates, pregnancy outcomes and implantation rates. The mean age (±SE) of PGS patients was 35.1± 1.6 (-HA, n=13) and 35.4±1.4 (+HA, n=17). μS-VTF resulted in over 99% survival of PGS-BLs independent of HA supplement. Furthermore, no difference (P≥ 0.2) in clinical pregnancy rates (+HA: 94.1%; -HA: 84.6%) or implantation rates (+HA: 87.5%; -HA: 72.0%) was determined to-date, eventhough the mean number of BLs transferred tended to be lower in the +HA group (1.33 vs 1.67). All single euploid BL-VETs (17 of 30) resulted in a pregnancy. In combination with ICE DMSO-free BL-VTF solutions, BL viability remains unchanged from its fresh-state. The addition of a large macromolecule like HA offers dual support by increasing both the viscosity of the VTF solution and supplemental cellular adhesion/membrane stabilization. If the trend toward higher implantation rates +HA becomes significant with a higher sample size, it would support the latter theory, as all BLs were cultured and transferred in the presence of HA. Overall, the success of μS-VTF of BLs has greatly facilitated single ET and Day5/6 trophectoderm biopsy/PGS as practical ART options.
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