Ferré L B, Chitwood J L, Fresno C, Ortega H H, Kjelland M E, Ross P J
Instituto Nacional de Tecnología Agropecuaria, Rafaela, Santa Fe, Argentina.
Department of Animal Science, University of California, Davis, CA, USA.
Reprod Domest Anim. 2018 Feb;53(1):26-33. doi: 10.1111/rda.13048. Epub 2017 Sep 11.
Straws of sex-sorted sperm are usually packaged at a low concentration (e.g., ~2.1 × 10 sperm/ml) and cost significantly more than unsorted conventional semen from the same sire. In order to maximize the efficiency of using sex-sorted sperm under in vitro fertilization conditions, the selection of an appropriate sperm separation technique is essential. In this study, the effect of using different silane-coated silica colloid dilutions and layering configurations during centrifugation of sex-sorted sperm was examined over an extended period of incubation time. Sperm recovery and viability after centrifugation using the colloid separation technique were measured along with several sperm motility parameters using CASA. For this purpose, frozen and thawed sex-sorted sperm samples were centrifuged using mini-volume single-layer (40%, 60% and 80%) and mini-volume two-layer (45%/90%, 40%/80% and 30%/60%) separation configurations using PureSperm . A single layer of 40% PureSperm recovered significantly more sex-sorted sperm (78.07% ± 2.28%) followed by a single layer of 80% PureSperm (68.43% ± 2.33%). The lowest sperm recovery was obtained using a two-layer PureSperm dilution of 45%/90% (47.57% ± 2.33%). Single-layer centrifugation recovered more sorted sperm (68.67% ± 1.74%) than two layer (53.74% ± 1.74%) (p < .0001). A single layer of 80% PureSperm exhibited the highest sorted sperm viability (72.01% ± 2.90%) after centrifugation (p < .05). The mini-volume single layer of 80% PureSperm was determined to be an effective alternative to a two-layer centrifugation configuration for sex-sorted sperm selection. In addition, single-layer colloid dilution of 80% performed either as well as or significantly outperformed the other treatments, as well as the control, with regard to motility (MOT) for all time periods of analysis.
性别分选精子细管通常以低浓度包装（例如，约2.1×10⁶精子/毫升），且成本比来自同一父本的未分选常规精液高得多。为了在体外受精条件下最大限度地提高使用性别分选精子的效率，选择合适的精子分离技术至关重要。在本研究中，在延长的孵育时间内，研究了在性别分选精子离心过程中使用不同的硅烷包被硅胶胶体稀释液和分层配置的效果。使用胶体分离技术离心后的精子回收率和活力，以及使用计算机辅助精子分析（CASA）测量的几个精子运动参数。为此，使用PureSperm对冷冻解冻的性别分选精子样本进行小体积单层（40%、60%和80%）和小体积双层（45%/90%、40%/80%和30%/60%）分离配置的离心。单层40%的PureSperm回收的性别分选精子显著更多（78.07%±2.28%），其次是单层80%的PureSperm（68.43%±2.33%）。使用45%/90%的双层PureSperm稀释液获得的精子回收率最低（47.57%±2.33%）。单层离心回收的分选精子（68.67%±1.74%）比双层（53.74%±1.74%）更多（p<.0001）。单层80%的PureSperm在离心后表现出最高的分选精子活力（72.01%±2.90%）（p<.05）。对于性别分选精子选择，80%的小体积单层被确定为双层离心配置的有效替代方案。此外，在所有分析时间段内，80%的单层胶体稀释液在运动性（MOT）方面与其他处理以及对照表现相当或显著优于它们。
