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High hydrostatic pressure: a new way to improve in vitro developmental competence of porcine matured oocytes after vitrification

¹ Population Genetics and Embryology, Institute of Genetics and Biotechnology and² Institute of Human Genetics, University of Aarhus, DK-800 Aarhus, Denmark³ Clinic for Large Animals, Faculty of Veterinary Science, Szent István University, 2225 Üll, Dóra major, Hungary⁴ Beijing Genomics Institute, Airport-Industrial zone B 6#, Beijing 101300, China and⁵ Kato Ladies' Clinic, Nishishinjuku, Shinjuku, Tokyo 160-0023, Japan

Correspondence should be addressed to Y Du at Faculty of Agricultural Sciences, Institute of Genetics and Biotechnology, University of Aarhus, Blichers Alle 20, Postbox 50, 8830 Tjele, Denmark; Email: duyt{at}genomics.org.cn

The purpose of the present study was to improve cryotolerance using high hydrostatic pressure (HHP) pretreatment of porcine in vitro matured (IVM) oocytes, to facilitate their further developmental competence after parthenogenetic activation. A total of 1668 porcine IVM oocytes were used in our present study. The pressure tolerance and optimal duration of recovery after HHP treatment were determined. Oocytes were treated with either 20 or 40 MPa (200 and 400 times greater than atmospheric pressure) for 60 min, with an interval of 10, 70, and 130 min between pressure treatment and subsequent vitrification under each pressure parameter. Oocytes from all vitrification groups had much lower developmental competence than fresh oocytes (P<0.01) measured as cleavage and blastocyst rates. However, significantly higher blastocyst rates (P<0.01) were obtained in the groups of 20 MPa pressure, with either 70 (11.4±2.4%) or 130 (13.1±3.2%) min recovery, when compared with the vitrification control group without HHP treatment where no blastocysts were obtained. The influence of temperature at HHP treatment on further embryo development was also investigated. Treatments of 20 MPa with 70 min recovery were performed at 37 °C or 25 °C. Oocytes pressurized at 37 °C had a significantly higher blastocyst (14.1±1.4%) rate than those treated at 25 °C (5.3±1.1%; P<0.01). Our results demonstrate that HHP pretreatment could considerably improve the developmental competence of vitrified pig in vitro matured (IVM) oocytes. The HHP pretreatment will be tested as a means to improve survival and developmental competence at different developmental stages in different species including humans.

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