Solid‐Surface Vitrification and In‐Straw Dilution After Warming of In Vitro‐Produced Bovine Embryos. Issue 1 (28th August 2013)

• Record Type: Journal Article
• Authors: Rodriguez‐Villamil, P; Ongaratto, FL; Fernandez Taranco, M; Bó, GA
• Is Part Of: Reproduction in domestic animals. Volume 49:Issue 1(2014)
• Page Start: 79
• Page End: 84
• Publication Date: 2013-08-28
• Subjects: Animal breeding -- Periodicals; Veterinary pathology -- Periodicals; Livestock -- Breeding -- Periodicals; 636.082
• DOI: 10.1111/rda.12229
• Languages: English
• ISSNs: 0936-6768

Abstract

<abstract abstract-type="main" id="rda12229-abs-0001"> <title>Contents</title> <p>Three experiments were designed to test a solid‐surface vitrification system for bovine <italic>in vitro</italic>‐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: VS1 (20% ethylene glycol (EG) + 20% propanediol (PROH) + 0.25 <sc>m</sc> trehalose (Tr)), VS2 (20% EG + 1M Tr) or VS3 (30% EG + 0.75 <sc>m</sc> Tr). Re‐expansion and hatching rates were higher (p < 0.05) for embryos vitrified in VS3 (72.2 ± 1.9 and 58.2 ± 0.8) than VS1 (64.4 ± 0.9 and 37.2 ± 2.5) or VS2 (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 VS1 or VS3 solutions. No significant differences were detected between the two methods; but re‐expansion and hatching rates were higher (p < 0.05) with VS3 (73.5 ± 3.1 and 47.1 ± 2.1) than VS1 (63.3 ± 3.3 and 39.7 ± 2.8). In experiment 3, embryos were vitrified by solid surface in VS1 or VS3 solutions and cryoprotectants were diluted in‐straw after warming in a TCM 199, 0.25 <sc>m</sc> sucrose solution or holding media. Survival rates of embryos vitrified in VS3 did not differ between those exposed to 0.25 <sc>m</sc> 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 VS1 were higher (p < 0.05) in those exposed to 0.25 <sc>m</sc> 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 <italic>in vitro</italic>‐produced bovine embryos.</p> </abstract>