A modified swim-up method reduces polyspermy during in vitro fertilization of porcine oocytes
A modified swim-up method reduces polyspermy during in vitro fertilization of porcine oocytes
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DOI：
10.1016/j.anireprosci.2008.12.004
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发表时间：
2009-10-01
影响因子：
2.2
通讯作者：
Lee, Chang-Kyu
中科院分区：
文献类型：
--
作者：
Park, Chi-Hun;Lee, Sang-Goo;Lee, Chang-Kyu
猪体外受精（IVF）的一般方法，涉及两个配子在培养基中共同培养，被认为是多精发生率高的主要原因。本研究的目的是在普通精子游动技术的基础上，采用改良的游动法减少体外受精过程中猪胚胎的多精受精。在这个设计中，一个70 pm孔径的细胞过滤器被用来分离放置在试管底部的精子和放置在上部区域的成熟卵母细胞。使用这种可渗透屏障分离配子是为了确保只有活动的精子才能进入卵母细胞。结果表明，与普通微滴法相比，改良游动法配制的3个种猪精子的多精率显著降低（p < 0.05）。然而，发现两种方法对两个品种的猪的渗透率是相似的。改良游泳法的囊胚平均出现率较高，总细胞数较多，但两种体外受精方法的囊胚率无显著差异。改良游动法的正常二倍体胚率显著高于微滴法（p < 0.05），而多倍体胚率显著高于改良游动法（p < 0.05）。结果表明，改良的游泳试管婴儿方法可以减少多精子的渗透，从而在猪体外受精中获得更好的质量和核型正常的胚胎。(C) 2008 Elsevier B.V.版权所有
The general method of porcine in vitro fertilization (IVF), involving the co-culture of both gametes in a medium drop, is thought to be the main reason for the high incidence of polyspermy. The aim of this study was to reduce the polyspermic fertilization of porcine embryos during IVF by the modified swim-up method, based on general sperm swim-up technique. Within this design, a 70 pm pore sized cell strainer was used to separate the sperm pellet placed at the bottom of a tube from the mature oocytes placed within the upper region. The separation of gametes using this permeable barrier was to ensure that only motile sperm gained access to the oocytes. It was found that the rate of polyspermy was significantly lowered for the sperm preparations from three boar breeds in modified swim-up method when compared with that of the general microdrop method (p < 0.05). However, the penetration rates were found to be similar in both methods for two boar breeds. The average occurrence of blastocysts with more total cell number was higher in the modified swim-up method, while no significant difference in blastocyst rates between the two IVF methods was observed. The frequency of normal diploid embryos was also significantly higher in the modified swim-up method and polyploidy was more frequently observed in microdrop method (p < 0.05). Our results demonstrated that the modified swim-up IVF method could reduce polyspermic penetration, and consequently produce better quality and karyotypically normal embryos in porcine IVF. (C) 2008 Elsevier B.V. All rights reserved.