使用流式细胞仪分离精子进行仔猪性别控制的研究

本研究旨在探索流式细胞仪分离精子在猪性别控制中的应用。使用流式细胞仪分离猪XY精子,而后通过母猪输卵管授精生产"预知"性别的仔猪,并使用吖啶橙染色法检测粗分离对精子核酸含量的影响。结果,成功利用分离获得的猪X和Y精子对母猪进行输卵管授精,母猪怀孕率、产仔率均为100%;输Y精子母猪产仔雄性率100%(♂6/6),对照母猪产仔雄性率57.14%(♂8/14);3头输X精子母猪产母仔率91.67%(♀11/12),对照母猪产雌性仔猪40%(♀2/5);使用性别分离精子不影响母猪的怀孕率、产仔率,但窝产仔数较低;吖啶橙染色法检测结果表明,流式细胞仪粗分离对猪精子核酸含量没有显著影响(P>0.05)。本研究结果提示,使用流式细胞仪分离精子授精可以有效改变仔猪的性别比例。本研究结果为猪分离精子性别控制技术的推广应用奠定了基础。

Sex-preselected Piglets Derived from Sexed Sperm by Flow Cytometry Sorting

The main objective of this study was to evaluate the efficiency of surgically artificial insemination(XY sperms were srperated by flow cytometry,and then the prevision sex piglets were breeded by artificial insemination) by using the sexed sperm sorted by flow cytometry to produce sex-preselected piglet.The effect of the sorting procedure on nucleic acid content of sperm was analyzed by acridine orange(AO) staining.Three recipient sows were inseminated with sorted X-sperm and were all pregnant and gave birth to a total of 12 piglets,from which 11 were females representing a 91.67% sexing accuracy,while 40% of piglets were female in the control group that inseminated with unsexed sperm.One sow inseminated with sorted Y-sperm farrowed 6 male piglets(100%,6/6),while 57.14% of male piglets in the control group.The total of sexing accuracy in this study was 94.4%(17/18) and no abnormality was observed in piglets derived from sexed sperm comparing to those derived from unsorted sperm.There was also no significant difference in pregnancy and farrowing rates between sows inseminated with sexed and unsexed sperm,but the litter size of sows inseminated with sexed sperm were lower than that of sows inseminated with unsexed sperm.There was no significant difference in the nucleic acid content between unsorted sperm and bulk sorted sperm(P>0.05).Results of this study indicate the feasibility of piglet production from sperm sorted by flow cytometry.