摘要
This study was conducted to develop an intrauterine inseminator (IUI) to deposit of frozen semen into uterus and to evaluate the results obtained after artificial insemination by IUI. Two Japanese spitzs (2 to 4 years of age) were used as semen donors. Semen was collected by manual masturbation into sterile glass collection tubes and separated into 3 fractions with only the sperm-rich fractions retained for further examination. Sperm motility >70%, sperm concentration of 200 to $400{\times}10^6 cells/ml$$\times$g for 5 min and poured out the suspended solution, and then diluted with 2 ml Tris-buffer which was consisted of 2.4 g Tris, 1.4 g citric acid, 0.8 g glucose, 0.1 $\mu\textrm{g}$/ml streptomycin, 100 IU/ml penicillin, 20 ml egg yolk to 100 ml mili-Q water (Ext I) or supplemented with 8 ml glycerol and 1 ml Equex STM paste to 100 rnl (Ext II). The diluted semen was cooled to 5$^{\circ}C$ in cold room, where the temperature in the sample reached 5$^{\circ}C$. Two h after beginning the cooling procedure, 2 ml of Ext II, also at 5$^{\circ}C$, was added and mixed by gently reversing the tubes several times during 1 h. The final sperm concentration for freezing was approximately $50{\times}10^6 cells/ml$. After equilibration, the semen was loaded into 0.5 ml straw and frozen on the liquid nitrogen vapour in styrofoam box. The straws were thawed at 7$0^{\circ}C$ for precisely 6 sec. After thawing of each straw, the frozen semen can survived over 50% motility. All the females were inseminated twice with 1 ml of $25{\times}10^6 cells/ml$
摘要译文
进行这项研究的目的是开发一种宫内授精器（IUI），以将冷冻的精液沉积到子宫中，并评估由IUI人工授精后获得的结果。使用了两个日本波茨坦犬（2-4岁）作为精液捐献者。通过手动手淫将精液收集到无菌玻璃收集管中，并分成3个馏分，仅保留富含精子的馏分以供进一步检查。精子活力> 70％，精子浓度为200至$ 400 {-次} 10 ^ 6个细胞/ ml $$-g，持续5分钟，倒出悬浮液，然后用2 ml Tris-缓冲液稀释取2.4克Tris，1.4克柠檬酸，0.8克葡萄糖，0.1美元/亩链霉菌素/毫升链霉素，100 IU /毫升青霉素，20毫升蛋黄至100毫升mili-Q水（Ext I）或补充8 ml甘油和1 ml Equex STM糊剂至100 mL（Ext II）。将稀释后的精液在冷室中冷却至5℃，样品中的温度达到5℃。在开始冷却程序两小时后，加入2 ml的Ext II（也为5 $ C），在1小时内轻轻颠倒试管几次，进行混合。冷冻的最终精子浓度约为$ 50 {-次} 10 ^ 6细胞/ ml $。平衡后，将精液装入0.5 ml秸秆中，并在泡沫聚苯乙烯盒中的液氮蒸气上冷冻。将秸秆在7 $ 0 ^ {circ} C $的温度下融化6秒钟。将每根稻草融化后，冷冻的精液可以存活超过50％的运动力。用1 ml的$ 25 {-次} 10 ^ 6细胞/ ml $对所有雌性进行了两次授精。
공일근조성균임용택이상인위성하. Production of Pups Following Artificial Insemination by Canine Intrauterine Inseminator[J]. Journal of Veterinary Clinics 한국임상수의학회지, 1999,16(2): 375-380