Transvaginal ultrasound-guided ovum pick up (OPU) has been widely utilized as a noninvasive and reliable technique that enables repeated retrieval of oocytes from live animals. To our knowledge, however, there are few reports of attempts to improve the follicle aspiration system for OPU in pigs. In this study, we examined the effects of different aspiration vacuum pressures and the phases of estrous cycle on oocyte recovery, the morphology of cumulus oocyte complexes (COCs), and blastocyst formation in Berkshire pigs. Firstly, three different vacuum pressures were used (60, 90 and 120 mmHg) and retrieved oocytes were classified based on their morphology of cumulus cells. There were no differences in the oocyte recovery rate among the aspiration vacuum pressures (5.3, 7.1 and 5.8 oocytes/head, respectively), however, the proportion of grade A oocytes (oocytes with several compact cumulus layers) in 90 mmHg (27.2%) was significantly higher (p<0.01) than in 120 mmHg (5.2%). Secondly, three phases of the estrous cycle (Day 2-5, Day 6-10 and Day 11-16; the final day of estrus was designed as Day 0 of the estrous cycle) were examined and there were no differences in the oocyte recovery rate among the estrous cycle phases (6.8, 6.3 and 8.3 oocytes/head, respectively). When all retrieved COCs were subjected to
in vitro maturation-
in vitro fertilization-
in vitro culture, there were no differences in cleavage and blastocyst formation rates per oocyte retrieval in the estrous cycle phase, and blastocysts were produced by retrieved COCs from every stage of estrous. These results show that grade A oocytes are effectively retrieved with 90 mmHg of aspiration vacuum pressure and that application of the OPU technique enables us repeatedly to obtain embryos from highly valuable live pigs.
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