title: Successful production of piglets derived from expanded blastocysts vitrified using a micro volume air cooling method without direct exposure to liquid nitrogen.
abstract: This study was conducted to clarify the feasibility of newly developed vitrification techniques for porcine embryos using the micro volume air cooling (MVAC) method without direct contact with liquid nitrogen (LN₂). Expanded blastocysts were vitrified in a solution containing 6 M ethylene glycol, 0.6 M trehalose and 2% (wt/vol) polyethylene glycol in 10% HEPES-buffered PZM-5. The blastocysts were collected from gilts and vitrified using the new device (MVAC) or a Cryotop (CT). Blastocysts were stored in LN₂ for at least 1 month. After warming, cryoprotective agents were removed using a single step. Survival of the embryos was assessed by in vitro culture (Experiment 1) and by embryo transfer to recipients (Experiment 2). In Experiment 1, the embryos vitrified by the MVAC or CT and fresh embryos without vitrification (Control) were used. The survival rates of embryos in the MVAC, CT and Control groups were 88.9% (32/36), 91.7% (33/36) and 100% (34/34), respectively, after 48 h culture, and the hatching rates of embryos after 48 h incubation were 69.4% (25/36), 63.9% (23/36) and 94.1% (32/34), respectively. In Experiment 2, 64 vitrified embryos were transferred to 5 recipient gilts, and 8 healthy piglets were produced from 3 recipients in the MVAC group. Similarly, 66 vitrified embryos were transferred to 5 recipient gilts, and 9 healthy piglets were produced from 2 recipients in the CT group. These results indicated that porcine expanded blastocysts can be cryopreserved using the MVAC method without potential pathogen contamination from LN₂.
authors: National Livestock Breeding Center, Fukushima 961-8511, Japan.
authors: Koji
authors: K
authors: Misumi
authors: Yuri
authors: Y
authors: Hirayama
authors: Sachiko
authors: S
authors: Egawa
authors: Shoko
authors: S
authors: Yamashita
authors: Hiroyoshi
authors: H
authors: Hoshi
authors: Kei
authors: K
authors: Imai
mesh_terms: D000818:Animals; D017849:Animals, Inbred Strains; D001755:Blastocyst / Q000187:drug effects; D003433:Crosses, Genetic; D015925:Cryopreservation / Q000295:instrumentation / Q000379:methods / Q000662:veterinary*; D003451:Cryoprotective Agents / Q000494:pharmacology; D004477:Ectogenesis / Q000187:drug effects; D046149:Embryo Culture Techniques / Q000662:veterinary; D010064:Embryo Implantation / Q000187:drug effects; D004624:Embryo Transfer / Q000662:veterinary*; D019855:Ethylene Glycol / Q000494:pharmacology; D005240:Feasibility Studies; D005260:Female; D007315:Insemination, Artificial / Q000662:veterinary; D007564:Japan; D050498:Live Birth / Q000662:veterinary; D008297:Male; D011092:Polyethylene Glycols / Q000494:pharmacology; D011247:Pregnancy; D034421:Sus scrofa / Q000502:physiology*; D014199:Trehalose / Q000494:pharmacology; D058989:Vitrification
chemical_list: D003451:Cryoprotective Agents; D011092:Polyethylene Glycols; D014199:Trehalose; D019855:Ethylene Glycol
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