Brüssow K P, Torner H, Rátky J, Hunter M G, Nürnberg G
Research Institute for the Biology of Farm Animals, Dummerstorf, Germany.
Acta Vet Hung. 1997;45(2):189-96.
Endoscopical ovum pick up (OPU) in swine is a minimal invasive and reliable technique to aspirate oocytes from preovulatory follicles for studying intrafollicular development and oocyte maturation as well as for IVM/IVF-programs. The aim of the present study was to determine the effect of different aspiration vacuum pressures on oocyte recovery and on the morphology of cumulus oocyte complexes (COCs). Oestrus of crossbred Landrace gilts (n = 33) was synchronized by feeding altrenogest and follicular growth was stimulated with 1,000 IU PMSG 24 h after the last altrenogest application. On day 4 after PMSG application preovulatory healthy follicles of > 5 mm diameter were aspirated laparoscopically. Aspiration was carried out using a two-way cannula and an electronic aspiration pump. Five different vacuum pressures were used: 10, 17, 32, 47 and 66 ml water/min, corresponding to 30, 60, 125, 250 and 375 mm Hg, respectively. Fluids from different follicles were pooled per ovary and the morphology of cumulus oocyte complexes (COCs) was determined microscopically immediately after aspiration. COCs were classified into oocytes with intact cumulus (i.e. compact or slightly expanded COC), oocytes with corona radiata and denuded oocytes. A total of 695 follicles were aspirated and 501 COCs recovered. Increasing the aspiration pressure stepwise from 10 ml water/min up to 66 ml water/min resulted in a decrease in oocyte recovery rate. A significant higher (P < 0.05) rate of oocyte recovery (77.4% v.s. 59.8%) was achieved using a vacuum pressure of 17 ml water/min compared to 66 ml water/min, respectively. There was a tendency to reduce the portion of COCs with intact cumulus from 82 to 88% to 77% if the vacuum pressure increased to more than 47 ml water/min. A higher aspiration pressure provoked an increase (P < 0.05) in the number of denuded oocytes: 0 to 3% at 10 to 32 ml water/min, respectively, compared to 10% at 47 ml water/min to 17% at 66 ml water/min. These results demonstrate that variation in aspiration pressure affects oocyte recovery rate and COC quality. Aspiration vacuum pressure of 17 to 32 ml water/min was found to be optimal in swine both for oocyte recovery and COC quality.
猪的内窥镜取卵(OPU)是一种微创且可靠的技术,可从排卵前卵泡中抽吸卵母细胞,用于研究卵泡内发育和卵母细胞成熟,以及用于体外成熟/体外受精程序。本研究的目的是确定不同抽吸真空压力对卵母细胞回收以及卵丘卵母细胞复合体(COC)形态的影响。通过饲喂烯丙孕素使杂交长白母猪(n = 33)发情同步,并在最后一次施用烯丙孕素24小时后用1000 IU孕马血清促性腺激素刺激卵泡生长。在施用孕马血清促性腺激素后的第4天,通过腹腔镜抽吸直径大于5 mm的排卵前健康卵泡。使用双向套管和电子抽吸泵进行抽吸。使用了五种不同的真空压力:10、17、32、47和66毫升水/分钟,分别对应于30、60、125、250和375毫米汞柱。每个卵巢将来自不同卵泡的液体汇集在一起,并在抽吸后立即在显微镜下确定卵丘卵母细胞复合体(COC)的形态。COC被分为卵丘完整的卵母细胞(即紧密或轻度扩张的COC)、有放射冠的卵母细胞和裸卵。总共抽吸了695个卵泡,回收了501个COC。将抽吸压力从10毫升水/分钟逐步增加到66毫升水/分钟会导致卵母细胞回收率降低。与66毫升水/分钟相比,使用17毫升水/分钟的真空压力分别实现了显著更高(P < 0.05)的卵母细胞回收率(77.4%对59.8%)。如果真空压力增加到超过47毫升水/分钟,有将卵丘完整的COC比例从82%至88%降低到77%的趋势。较高的抽吸压力会导致裸卵数量增加(P < 0.05):在10至32毫升水/分钟时分别为0%至3%,而在47毫升水/分钟时为10%,在66毫升水/分钟时为17%。这些结果表明,抽吸压力的变化会影响卵母细胞回收率和COC质量。发现17至32毫升水/分钟的抽吸真空压力对猪的卵母细胞回收和COC质量均为最佳。