Parrilla Inmaculada, Vazquez Juan M, Gil Maria A, Caballero Ignacio, Almiñana Carmen, Roca Jordi, Martinez Emilio A
Department of Animal Medicine and Surgery, Faculty of Veterinary, University of Murcia, 30071 Murcia, Spain.
Theriogenology. 2005 Jul 1;64(1):86-98. doi: 10.1016/j.theriogenology.2004.11.004. Epub 2004 Dec 15.
Sex-sorting of boar spermatozoa is an emerging biotechnology, still considered suboptimal owing to the slowness of the process, which requires long sorting periods to obtain an adequate number of spermatozoa to perform a non-surgical insemination. This period involves storage of sorted cells that could impair their functional capacity. Here, we have studied how the storage of sex-sorted boar spermatozoa affects their functional capacity. Sorted spermatozoa were assessed at various times (0, 2, 5h or 10h) during storage after sorting and compared with diluted and unsorted spermatozoa for sperm motility patterns, plasma membrane and acrosomal integrity and their ability to penetrate homologous IVM oocytes. Sex-sorted sperm motility and membrane integrity only decreased significantly (p<0.05) by the end of the storage period (10h) compared to unsorted spermatozoa. Sperm velocity, ALH and Dance increased significantly (p<0.05), immediately post-sorting, returning to unsorted sperm values during storage. Acrosome integrity was not seriously affected by the sorting process, but decreased (p<0.05) during storage after sorting. Sorted spermatozoa stored 2h after sorting did not differ from unsorted in penetration rates and numbers of spermatozoa per oocyte, reaching the highest (p<0.05) penetration rates and sperm numbers per oocyte, when co-cultured for 6 or more hours. Non-storage or storage for 5h or 10h negatively (p<0.05) affected sperm penetration ability. In conclusion, although flow cytometrically sex-sorted spermatozoa are able to maintain motility, viability and acrosomal integrity at optimal levels until 10h of storage after sorting, fertilizing ability is maintained only over shorter storage times (<5h).
公猪精子的性别分选是一项新兴的生物技术，由于该过程缓慢，仍被认为不够理想，这需要较长的分选时间才能获得足够数量的精子来进行非手术授精。这段时间涉及对分选后的细胞进行储存，这可能会损害它们的功能能力。在此，我们研究了性别分选的公猪精子储存对其功能能力的影响。在分选后的储存过程中的不同时间点（0、2、5小时或10小时）对分选后的精子进行评估，并与稀释的未分选精子在精子活力模式、质膜和顶体完整性以及穿透同源体外成熟卵母细胞的能力方面进行比较。与未分选的精子相比，性别分选的精子活力和膜完整性仅在储存期结束时（10小时）显著下降（p<0.05）。分选后立即精子速度、平均路径速度和鞭打频率显著增加（p<0.05），在储存期间恢复到未分选精子的值。顶体完整性不受分选过程的严重影响，但在分选后的储存过程中下降（p<0.05）。分选后储存2小时的分选精子在穿透率和每个卵母细胞的精子数量方面与未分选的精子没有差异，当共同培养6小时或更长时间时，达到最高（p<0.05）的穿透率和每个卵母细胞的精子数量。不储存或储存5小时或10小时会对精子穿透能力产生负面影响（p<0.05）。总之，尽管通过流式细胞术进行性别分选的精子能够在分选后长达10小时的储存时间内将活力、生存力和顶体完整性维持在最佳水平，但受精能力仅在较短的储存时间（<5小时）内得以维持。
