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Comparison of vitrification and conventional freezing for cryopreservation of caprine embryos

  • Paula F.B. Araújo-Lemos (a1), Leopoldo M. Freitas Neto (a2), Marcelo T. Moura (a2), Janaína V. Melo (a3), Paulo F. Lima (a2) and Marcos A.L. Oliveira (a4)...


The experiment aimed to compare conventional freezing and different vitrification protocols for cryopreservation of caprine embryos at morphological, ultrastructural, and functional levels. Caprine embryos produced in vivo were allocated randomly to three groups: (1) conventional freezing with ethylene glycol (EG); (2) dimethyl sulfoxide + EG (DMSO/EG) vitrification; and (3) dimethylformamide + EG (DMF/EG) vitrification. All groups were scored for cell viability (propidium iodide staining and ultrastructural levels) and re-expansion rate after thawing or warming. Embryos subjected to DMSO/EG vitrification showed higher cell viability (73.33%), compared with DMF/EG vitrification and conventional freezing group embryos (40.00 and 66.66%, respectively). The ultrastructural study revealed that vitrified embryos had greater preservation of cellular structure than embryos from conventional freezing with EG. DMSO/EG vitrification resulted in higher rates of re-expansion in vitro (47.36%) than DMF/EG vitrification (31.58%), and conventional freezing (25.00%). In conclusion, caprine embryos produced in vivo are better cryopreserved after vitrification than conventional freezing, therefore we conclude that DMSO/EG vitrification is the most effective protocol for cryopreservation.


Corresponding author

All correspondence to: Marcos A.L. de Oliveira. Laboratório de Biotécnicas Aplicadas a Reprodução Animal, Departamento de Medicina Veterinária, Universidade Federal Rural de Pernambuco (UFRPE), Rua Dom Manoel de Medeiros s/n, Dois Irmãos, CEP 52171–900, Recife-PE, Brasil. e-mail:


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Al Yacoub, A.N., Gauly, M. & Holtz, W. (2010). Open pulled straw vitrification of goat embryos at various stages of development. Theriogenology 73, 1018–23.
Alvarenga, M.A., Papa, F.O., Landim-Alvarenga, F.C. & Medeiros, A.S. (2005). Amides as cryoprotectants for freezing stallion semen: a review. Anim. Reprod. Sci. 89, 105–13.
Bezerra, F.S., Castelo, T.S., Alves, H.M., Oliveira, I.R., Lima, G.L., Peixoto, G.C., Bezerra, A.C. & Silva, A.R. (2011). Objective assessment of the cryoprotective effects of dimethylformamide for freezing goat semen. Cryobiology 63, 263–6.
Bilton, R.J. & Moore, N.W. (1976). In vitro culture, storage and transfer of goat embryos. Aust. J. Biol. Sci. 29, 125–9.
Castro, S.V., de Carvalho, A.A., da Silva, C.M., Faustino, L.R., Campello, C.C., Lucci, C.M., Báo, S.N., de Figueiredo, J.R. & Rodrigues, A.P. (2011). Freezing solution containing dimethylsulfoxide and fetal calf serum maintains survival and ultrastructure of goat preantral follicles after cryopreservation and in vitro culture of ovarian tissue. Cell Tissue Res. 346, 283–92.
Chalah, T., Seigneurin, F., Blesbois, E. & Brillard, J.P. (1999). In vitro comparison of fowl sperm viability in ejaculates frozen by three different techniques and relationship with subsequent fertility in vivo. Cryobiology 39, 185–91.
Chen, S.L. & Tian, , , Y.S. (2005). Cryopreservation of flounder (Paralichthys olivaceus) embryos by vitrification. Theriogenology 63, 1207–19.
Cognié, Y., Baril, G., Poulin, N. & Mermillod, P. (2003). Current status of embryos technologies in sheep and goat. Theriogenology 59, 171–88.
Dike, I.P. (2009). Efficiency of intracellular cryoprotectants on the cryopreservation of sheep oocytes by controlled slow freezing and vitrification techniques. J. Cell Anim. Biol. 3, 44–9.
Dobrinsky, J.R. (2002). Advancements in cryopreservation of domestic animal embryos. Theriogenology 57, 285302.
El-Gayar, M. & Holtz, W. (2001). Technical note: Vitrification of goat embryos by the open pulled-straw method. J. Anim. Sci. 79, 2436–8.
Gibb, Z., Morris, L.H., Maxwell, W.M. & Grupen, C.G. (2013). Dimethyl formamide improves the postthaw characteristics of sex-sorted and nonsorted stallion sperm. Theriogenology 79, 1027–33.
Guignot, F., Bouttier, A., Baril, G., Salvetti, P., Pignon, P., Beckers, J.F., Touzé, J.L., Cognié, J., Traldi, A.S., Cognié, Y. & Mermillod, P. (2006). Improved vitrification method allowing direct transfer of goat embryos. Theriogenology 66, 1004–11.
Hong, Q.H., Tian, S.J., Zhu, S.E., Feng, J.Z., Yan, C.L., Zhao, X.M., Liu, G.S. & Zheng, S.M. (2007). Vitrification of Boer goat morulae and early blastocysts by straw and open-pulled straw method. Reprod. Domest. Anim. 42, 34–8.
Kasai, M., Niwa, K. & Iritani, A. (1981). Effects of various cryoprotective agents on the survival of unfrozen and frozen mouse embryos. J. Reprod. Fertil. 63, 175–80.
Kuleshova, L.L, Shaw, J.M. & Trounson, A.O. (2001). Studies on replacing most of the penetrating cryoprotectant by polymers for embryo cryopreservation. Cryobiology 43, 2131.
Loutradi, K.E., Kolibianakis, E.M., Venetis, C.A., Papanikolaou, E.G., Pados, G., Bontis, I. & Tarlatzis, B.C. (2008). Cryopreservation of human embryos by vitrification or slow freezing: a systematic review and meta-analysis. Fertil. Steril. 90, 186–93.
Lukaszewicz, E. (2002). An effective method for freezing White Italian gander semen. Theriogenology 58, 1927.
Malo, C., Gil, L., Cano, R., Martínez, F., García, A. & Jerez, R.A. (2012). Dimethylformamide is not better than glycerol for cryopreservation of boar semen. Andrologia 44(Suppl. 1), 605–10.
Massip, A. (1989). Some significant steps in the cryopreservation of mammalian embryos with a note on a vitrification procedure. Anim. Reprod. Sci. 19, 117–29.
Massip, A. (2001). Cryopreservation of embryos of farm animals. Reprod. Domest. Anim. 36, 4955.
Morató, R., Romaguera, R., Izquierdo, D., Paramio, M.T. & Mogas, T. (2011). Vitrification of in vitro produced goat blastocysts: effects of oocyte donor age and development stage. Cryobiology. 63, 240–4.
Moustacas, V.S., Cruz, B.C., Varago, F C., Miranda, D.A., Lage, P.G. & Henry, M. (2011). Extenders containing dimethylformamide associated or not with glycerol are ineffective for ovine sperm cryopreservation. Reprod. Domest. Anim. 46, 924–5.
Polge, C. (1951). Functional survival of fowl spermatozoa after freezing at –79°C. Nature 167, 949–50.
Stringfellow, D.A. & Seidel, S.M. (1998). Manual of the International Embryo Transfer Society: a procedural guide and general information for the use of embryo transfer technology emphasizing sanitary procedures, 3rd edn.Savory III, USA: International Embryo Transfer Society.
Vajta, G. (2000). Vitrification of oocytes and embryos of domestic animals. Anim. Reprod. Sci. 60–61, 357–64.
Vajta, G. & Kuwayama, M. (2006). Improving cryopreservation systems. Theriogenology 65, 236–44.
Whittingham, D.G., Leibo, S.P. & Mazur, P. (1972). Survival of mouse embryos frozen to –196°C and –296°C. Science 178, 411–4.
Wilmut, I. (1972). The effect of cooling rate, cryoprotective agent and stage of development on survival of mouse embryos during freezing and thawing. Life Sci. II 11, 1071–9.



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