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Post-thaw culture in presence of insulin-like growth factor I improves the quality of cattle cryopreserved embryos

  • Alexander V. Makarevich (a1), Elena Kubovičová (a2), Zdena Hegedušová (a2) (a3), Juraj Pivko (a2) and František Louda (a2)...

Summary

The goal of this study was to examine the effect of insulin-like growth factor I (IGF-I; added during post-thaw culture (48 h)) on the preimplantation viability and quality of cryopreserved bovine in vivo recovered embryos. The morula stage embryos, non-surgically recovered from superovulated dairy cows of Czech Fleckvieh cattle breed, had previously been cryopreserved by a slow freezing technique and stored in liquid nitrogen since 1989–1990. Following thawing, the embryos were cultured for 48 h either alone (no IGF-I) or in the presence of IGF-I (10 or 100 ng/ml); non-cultured embryos served as a control. Thereafter, the embryos were analyzed for cleavage to the blastocyst stage, apoptosis (TUNEL), embryo cell number and quality of actin cytoskeleton. Following post-thaw culture 41% of embryos developed to advanced blastocysts. IGF-I increased this per cent and, at a higher dose, essentially reduced the per cent of degenerated embryos. In cultured embryos, IGF-I at both doses elevated the cell number compared with non-cultured embryos. However, in comparison with embryos cultured without IGF-I, only the higher IGF-I dose resulted in elevating the embryo cell number. The TUNEL index was significantly lowered by IGF-I treatment. Thawed embryos were mostly of the grade III actin type and fewer (12%) had grade II actin, whilst no grade I actin embryos were noted. The addition of IGF-I resulted in the appearance of grade I actin embryos (8.33 and 6.9% for 10 and 100 ng/ml, respectively). These observations indicate that the addition of IGF-I during post-thaw culture can improve the quality of bovine cryopreserved embryos.

Copyright

Corresponding author

All correspondence to: Alexander V. Makarevich. Institute for Cattle Breeding Ltd Rapotin, Vyzkumniku 267, 788 13 Vikyrovice, Czech Republic. Tel.: +421 37 6546 334. Fax: +421 37 6546 480. e-mail: makarevic@cvzv.sk

References

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Block, J. (2007). Use of insulin-like growth factor-1 to improve post-transfer survival of bovine embryos produced in vitro. Theriogenology 68S, S49S55.
Block, J., Wrenzycki, C., Niemann, H., Herrmann, D. & Hansen, P.J. (2008). Effects of insulin-like growth factor-1 on cellular and molecular characteristics of bovine blastocysts produced in vitro. Mol. Reprod. Dev. 75, 895903.
Byrne, A.T., Southgate, J., Brison, D.R. & Leese, H.J. (1999). Analysis of apoptosis in the preimplantation bovine embryo using TUNEL. J. Reprod. Fert. 117, 97105.
Byrne, A.T., Southgate, J., Brison, D.R. & Leese, H.J. (2002). Regulation of apoptosis in the bovine blastocyst by insulin and the insulin-like growth factor (IGF) superfamily. Mol. Reprod. Dev. 62, 489–95.
Dobrinsky, J.R. (1996). Cellular approach to cryopreservation of embryos. Theriogenology 45, 1726.
Fabian, D., Gjorett, J.O., Berthelot, F., Martinat-Botte, F. & Maddox-Hyttel, P. (2005). Ultrastructure and cell death of in vivo derived and vitrified porcine blastocysts. Mol. Reprod. Dev. 70, 155–65.
Greve, T., Avery, B. & Callesen, H. (1993). Viability of in vitro and in vivo produced bovine embryos. Reprod. Dom. Anim. 28, 164–9.
Hernandez-Fonseca, H.J., Sirisathien, S., Bosch, P., Cho, H.S., Lott, J.D., Hawkins, L.L., Hollet, R.B., Coley, S.L. & Brackett, B.G. (2002). Offspring resulting from direct transfer of cryopreserved bovine embryos produced in vitro in chemically defined media. Animal Reprod. Sci. 69, 151–8.
Makarevich, A. & Markkula, M. (2002) Apoptosis and proliferation potential of bovine embryos stimulated with insulin-like growth factor-I during in vitro maturation and culture. Biol. Reprod. 66, 386–92.
Makarevich, A.V., Chrenek, P., Olexikova, L., Popelkova, M., Turanova, Z., Ostro, A. & Pivko, J. (2008). Post-thaw survival, cell death and actin cytoskeleton in gene-microinjected rabbit embryos after vitrification. Theriogenology 70, 675–81.
Marquez-Alvarado, Y.C., Galina, C.S., Castilla, B., Leon, H. & Morena-Mendoza, N. (2004). Evidence of damage in cryopreserved and fresh bovine embryos using the Tunel technique. Reprod. Dom. Anim. 39, 141–5.
Markkula, M., Raty, M., Jauhiainen, L., Paranko, J., Raula, J. & Makarevich, M. (2001). Ratio of proliferating cell nuclear antigen-positive nuclei to total cell number is higher in day 7 than in day 8 vitrified in vitro-produced bovine embryos. Biol. Reprod. 65, 52–9.
Mori, M, Otoi, T. & Suzuki, T. (2002). Correlation between the cell number and diameter in bovine embryos. Reprod. Dom. Anim. 37, 181–4.
Popelkova, M., Chrenek, P., Pivko, J., Makarevič, A.V., Kubovičová, E. & Kačmarik, J. (2005). Survival and ultrastructure of gene-microinjected rabbit embryos after vitrification. Zygote 13, 283–93.
Popelkova, M., Turanova, Z., Koprdova, L, Ostro, A., Toporcerova, S., Makarevich, A.V. & Chrenek, P. (2009). Effect of vitrification technique and assisted hatching on rabbit embryo developmental rate. Zygote 17, 5761.
Quetglas, M.D., Coelho, L.A., Garcia, J.M., Oliveira Filho, E.B. & Esper, C.R. (2001). Effect of insulin-like growth factor-1 during in vitro oocyte maturation and in vitro culture of bovine embryos. Arquivo Brasileiro de Medicina Veterinaria e Zootechnia 53, 15.
Říha, J., Machatková, M. & Pavlok, A. (2002). Viability of fresh and frozen transferred IVP bovine embryos. Czech J. Anim. Sci. 47, 261–7.
Rizos, D., Ward, F., Boland, M.P. & Lonergan, P. (2001). Effect of culture system on the yield and quality of bovine blastocysts as assessed by survival after vitrification. Theriogenology 56, 116.
Sirisathien, S. & Brackett, B.G. (2003). TUNEL analyses of bovine blastocysts after culture with EGF and IGF-I. Molec. Reprod. Dev. 65, 51–6.
Tharasanit, T., Colenbrander, B. & Stout, T.A.E. (2005). Effect of cryopreservation on the cellular integrity of equine embryos. Reproduction 129, 789–98.
Velazquez, M.A., Zaraza, J., Oropeza, A., Webb, R. & Niemann, H. (2009). The role of IGF1 in the in vivo production of bovine embryos from superovulated donors. Reproduction 137, 161–80.
Wright, R. & Ellington, J. (1995). Morphological and physiological differences between in vivo- and in vitro-produced preimplantation embryos from livestock species. Theriogenology 44, 1167–89.

Keywords

Post-thaw culture in presence of insulin-like growth factor I improves the quality of cattle cryopreserved embryos

  • Alexander V. Makarevich (a1), Elena Kubovičová (a2), Zdena Hegedušová (a2) (a3), Juraj Pivko (a2) and František Louda (a2)...

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