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Cryopreservation of Porcine Embryos Derived from In Vitro-Matured Oocytes¹

Laboratory of Developmental Engineering,⁵ Department of Life Science, School of Agriculture, Meiji University, Kawasaki 214-8571, Japan Chiba Prefectural Animal Experimental Station,⁶ Chiba, 289-1113, Japan Kato Ladies' Clinic,⁷ Shinjuku, Tokyo, 160-0023, Japan

This study describes a cryopreservation method for porcine in vitro-produced (IVP) embryos using as a model parthenogenetic embryos derived from in vitro-matured (IVM) oocytes. IVP embryos at the expanded blastocyst stage were cryopreserved by vitrification using the minimum volume cooling (MVC) method and exhibited an embryo survival rate of 41.2%. Survival was then significantly improved (83.3%, P < 0.05) by decreasing the amount of cytoplasmic lipid droplets (delipation) prior to vitrification. IVP embryos at the 4-cell stage also survived cryopreservation when vitrified after delipation (survival rate, 36.0%), whereas post-thaw survival of nondelipated embryos was quite low (9.7%). Furthermore, it was demonstrated that porcine IVP morulae can be cryopreserved by vitrification following delipation by a noninvasive method (survival rate, 82.5%). These results clearly confirm that porcine embryos derived from IVM oocytes can be effectively cryopreserved with high embryo survival using the MVC method in conjunction with delipation.

¹ This work was supported by grant from the Japan Livestock Technology Association, PROBRAIN and the National Institute of Agrobiological Science.

² Correspondence: Hiroshi Nagashima, Laboratory of Developmental Engineering, Department of Life Science, School of Agriculture, Meiji University, 1-1-1 Higashimita, Tama, Kawasaki 214-8571, Japan. FAX: 81 44 934 7824; hnagas{at}isc.meiji.ac.jp

³ Current address: Laboratory of Anatomy and Embryology, Institute of Basic Medical Science, Tsukuba University, 1-1-1, Tennoudai, Tsukuba 305-8577, Japan

⁴ Current address: National Institute of Genetics, 1111 Yata, Mishima, Shizuoka, 411-8540, Japan

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