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Solid‐Surface Vitrification and In‐Straw Dilution After Warming of In Vitro‐Produced Bovine Embryos
Author(s) -
RodriguezVillamil P,
Ongaratto FL,
Fernandez Taranco M,
Bó GA
Publication year - 2014
Publication title -
reproduction in domestic animals
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.546
H-Index - 66
eISSN - 1439-0531
pISSN - 0936-6768
DOI - 10.1111/rda.12229
Subject(s) - vitrification , cryoprotectant , hatching , straw , zoology , sucrose , ethylene glycol , chemistry , biology , horticulture , botany , embryo , andrology , cryopreservation , food science , agronomy , medicine , fishery , organic chemistry
Contents Three experiments were designed to test a solid‐surface vitrification system for bovine in vitro ‐produced embryos and to develop a simple method of in‐straw dilution after warming, which can be potentially used for direct transfer in the field. Experiment 1 evaluated embryo survival rates (i.e. re‐expansion and hatching) after vitrification and warming in three different solutions: VS 1 (20% ethylene glycol ( EG ) + 20% propanediol ( PROH ) + 0.25 m trehalose (Tr)), VS 2 (20% EG + 1M Tr) or VS 3 (30% EG + 0.75 m Tr). Re‐expansion and hatching rates were higher (p < 0.05) for embryos vitrified in VS 3 (72.2 ± 1.9 and 58.2 ± 0.8) than VS 1 (64.4 ± 0.9 and 37.2 ± 2.5) or VS 2 (68.5 ± 1.5 and 49.6 ± 1.0; p < 0.05). Experiment 2 was designed to compare two methods of vitrification: glass micropipettes or solid surface, using the VS 1 or VS 3 solutions. No significant differences were detected between the two methods; but re‐expansion and hatching rates were higher (p < 0.05) with VS 3 (73.5 ± 3.1 and 47.1 ± 2.1) than VS 1 (63.3 ± 3.3 and 39.7 ± 2.8). In experiment 3, embryos were vitrified by solid surface in VS 1 or VS 3 solutions and cryoprotectants were diluted in‐straw after warming in a TCM 199, 0.25 m sucrose solution or holding media. Survival rates of embryos vitrified in VS 3 did not differ between those exposed to 0.25 m sucrose (74.7 ± 1.3 and 57.2 ± 2.2) or holding (77.3 ± 1.4 and 58.0 ± 2.5) medium after warming; however, survival rates of embryos vitrified in VS 1 were higher (p < 0.05) in those exposed to 0.25 m sucrose (67.7 ± 2.3 and 47.0 ± 1.7) than holding medium (54.5 ± 1.0 and 27.7 ± 3.1). In conclusion, solid‐surface vitrification using simplified EG ‐based solutions and in‐straw dilution with holding media may be a practical alternative for cryopreservation and direct transfer of in vitro ‐produced bovine embryos.