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| Production of Porcine (Sus scrofa) Embryos by ICSI Using Frozen Thawed Semen |
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Abstract Abstract Pig (Sus scrofa) is a good animal model for human disease and stem cell study, also is one of the important domestic species in the husbandry. In recent years, more and more scientific research needs pig preimplantation embryos for experiment material, due to the development and application of pig embryo engineering and related scientific research on porcine. Because of immaturation of the pig semen and embryo cryopreservation techniques, most of pig preimplantation embryos used in laboratory are obtained by in vitro fertilization (IVF) with fresh semen or frozen-thawed semen. Collection and transportation of fresh semen is inconvenient, and IVF with frozen semen is inefficient. Polyspermy is also easily to happen, which can lead to result mistake. Therefore, the establishment of a fast and efficient technological system for producing high-quality porcine embryos in vitro needs urgent resolve for scientists in the related research fields. In this study, we integrated and optimized porcine semen cryopreservation, oocyte in vitro maturation and oocyte intracytoplasmic injection (ICSI) techniques to establish a highly efficient pig embryo in vitro production system. First, we established stable system of oocyte in vitro maturation, the cleavage rate of parthenogenetic activation and blastocyst developmental rates were 79.51% and 32.30% respectively. And then, we performed IVF with fresh semen and frozen semen, embryo cleavage rates were 49.89% and 28.30% respectively, blastocyst developmental rates were 31.44% and 10.55% respectively. Embryo cleavage rate and blastocyst developmental rate of IVF with fresh semen were significantly higher than those of freeze-thawing semen (P<0.05). In addition, further test was conducted by in vitro production of pig embryo using the method of oocyte ICSI with frozen semen, the cleavage rate and blastocyst developmental rates were 83.00% and 23.32% respectively, while the embryo cleavage rate and blastocyst rates were 49.89% and 31.44% respectively by IVF with fresh semen. Blastocyst developmental rate of ICSI was lower than that of IVF and parthenogenetic activation. But for total blastocyst yield rate, it was significantly higher by oocyte ICSI than that of fresh semen IVF method. This study sets up an effective, economical system to produce embryos for scientific research and animal reproduction.
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Received: 27 December 2016
Published: 02 May 2017
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