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Effects of hyaluronic acid in culture and cytochalasin B treatment before freezing on survival of cryopreserved bovine embryos produced in vitro

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Summary

One limitation to the widespread use of in vitro-produced embryos in cattle is their poor survival following cryopreservation. Two approaches for enhancing survival of in vitro-produced bovine embryos following cryopreservation were evaluated: culture in the presence of hyaluronic acid and alterations in the cytoskeleton through cytochalasin B treatment. The experiment was a 2×2 factorial design to test main effects of hyaluronic acid added to culture at day 5 after insemination (+or−) and cryopreservation treatment (control or cytochalasin B). Embryos used for cryopreservation were blastocysts and expanded blastocysts harvested on day 7 after insemination. Cytochalasin B increased the percent of embryos that re-expanded (P<0.0001) and that hatched following thawing (P<0.05). The hatching percent was 29.6% for embryos treated with cytochalasin B versus 9.1% for control embryos. There was no significant effect of hyaluronic acid on survival although there was a tendency for embryos cultured with hyaluronic acid to have higher percent hatching if not treated with cytochalasin B (12.7% for hyaluronic acid versus 4.5% for control; hyaluronic acid x cytochalasin B interaction; P=0.09). In conclusion, cytochalasin B treatment before freezing improved cryosurvival of bovine embryos produced in vitro. Such a treatment could be incorporated into methods for cryopreservation of bovine embryos provided post-transfer survival is adequate. In contrast, culture with hyaluronic acid was of minimal benefit—the increased cryosurvival in the absence of cytochalasin B was not sufficient to allow an adequate number of embryos to survive.

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References

  • Abe, H.; Yamashita, S.; Itoh, T.; Satoh, T.; Hoshi, H. Ultrastructure of bovine embryos developed from in vitro-matured and fertilized oocytes: comparative morphological evaluation of embryos cultured either in serum-free medium or in serum supplemented medium. Mol. Reprod. Dev. 53:325–335; 1999.

    Article  PubMed  CAS  Google Scholar 

  • Agca, Y.; Monson, R. L.; Northey, D. L.; Abas Mazni, O.; Schaeffer, D. M.; Rutledge, J. J. Transfer of fresh and cryopreserved IVP bovine embryos: normal calving, birth weight and gestation lengths. Theriogenology 50:147–162; 1998.

    Article  PubMed  CAS  Google Scholar 

  • Al-Katanani, Y. M.; Drost, M.; Monson, R. L.; Rutledge, J. J.; Krininger, C. E., III; Block, J.; Thatcher, W. W.; Hansen, P. J. Pregnancy rates following timed embryo transfer with fresh or vitrified in vitro produced embryos in lactating dairy cows under heat stress condition. Theriogenology 58:171–182; 2002.

    Article  PubMed  CAS  Google Scholar 

  • Ambrose, J. D.; Drost, M.; Monson, R. L.; Rutledge, J. J.; Leifbfried-Rutledge, M. L. Efficacy of timed embryo transfer with fresh and frozen in vitro produced embryos to increase pregnancy rates in heat-stressed dairy cattle. J. Dairy Sci. 82:2369–2376; 1999.

    Article  PubMed  CAS  Google Scholar 

  • Bertolini, M.; Beam, S. W.; Shim, H.; Bertolini, L. R.; Moyer, A. L.; Famula, T. R.; Anderson, G. B. Growth, development, and gene expression by in vivo and in vitro produced day 7 and 16 bovine embryos. Mol. Reprod. Dev. 63:318–328; 2002.

    Article  PubMed  CAS  Google Scholar 

  • De La Torre-Sanchez J.; Gardner, D.; Preis, K.; Seidel, G., Jr. Regulation of glucose metabolism to decrease lipid content of in vitro-produced bovine embryos. Reprod. Fertil. Dev. 17:218 (abstract); 2005.

    Article  Google Scholar 

  • Diez, C.; Heyman, Y.; Le Bourhis, D.; Guyader-Joly, C.; Degrouard, J.; Renard, J. P. Delipidating in vitro-produced bovien zygotes: Effect on further development and consequences for freezability. Theriogenology 55:923–936; 2001.

    Article  PubMed  CAS  Google Scholar 

  • Dobrinsky, J. R. Cellular approach to cryopreservation of embryos. Theriogenology 45:17–26; 1996.

    Article  Google Scholar 

  • Dobrinsky, J. R.; Overström, E. W.; Duby, R. T.; Johnson, L. A.; Duffy, P.; Roche, J. F.; Boland, M. P. Effect of cytoskeletal stabilization on the development of bovine embryos cryopreserved by vitrification. Theriogenology 43:199 (abstract); 1995.

    Article  Google Scholar 

  • Dobrinsky, J. R.; Pursel, V. G.; Long, C. R.; Johnson, L. A. Birth of piglets after transfer of embryos cryopreserved by cytoskeletal stabilization and vitrification. Biol. Reprod. 62:564–570; 2000.

    Article  PubMed  CAS  Google Scholar 

  • Enright, B. P.; Lonergan, P.; Dinnyes, A.; Fair, T.; Ward, F. A.; Yang, X.; Boland, M. P. Culture of in vitro produced bovine zygotes in vitro vs in vivo: implications for early embryo development and quality. Theriogenology 54:659–673; 2000.

    Article  PubMed  CAS  Google Scholar 

  • Fair, T.; Lonergan, P.; Dinnyes, A.; Cotell, D. C.; Hyttel, P.; Ward, F. A.; Boland M. P. Ultrastructure of bovine blastocysts following cryopreservation: effects of method of blastocyst production. Mol. Reprod. Dev. 58:186–195; 2001.

    Article  PubMed  CAS  Google Scholar 

  • Fischer-Brown, A.; Monson, R.; Parrish, J.; Rutledge, J. Cell allocation in bovine embryos cultured in two media under two oxygen concentrations. Zygote 10:341–348; 2002.

    Article  PubMed  CAS  Google Scholar 

  • Furnus, C. C.; de Matos, D. G.; Martinez, A. G. Effect of hyaluronic acid on development of in vitro produced bovine embryos. Theriogenology 49:1489–1499; 1998.

    Article  PubMed  CAS  Google Scholar 

  • Guyader-Joly, C.; Ponchon, S.; Durand, M.; Heyman, Y.; Renard, J. P.; Ménézo, Y. Effect of lecithin on in vitro and in vivo survival of in vitro produced bovine blastocysts after cryopreservation. Theriogenology 52:1193–1202; 1999.

    Article  PubMed  CAS  Google Scholar 

  • Hansen, P. J.; Block, J. Towards an embryocentric world: the current and potential uses of embryo technologies in dairy production. Reprod. Fertil. Dev. 16:1–14; 2004.

    Article  PubMed  Google Scholar 

  • Hardingham, T. E.; Fosang, A. J. Proteoglycans: many forms and many functions. FASEB J. 6:861–870; 1992.

    PubMed  CAS  Google Scholar 

  • Hasler, J. F.; Henderson W. B.; Hurtgen, P. J., et al. Production, freezing and transfer of bovine IVF embryos and subsequent calving results. Theriogenology 43:141–152; 1995.

    Article  Google Scholar 

  • Hasler, J. F.; Hurtgen, P. J.; Jin, Z. Q.; Stokes, J. E. Survival of IVF-derived bovien embryos frozen in glycerol or ethylene glycol. Theriogenology 48:563–579; 1997.

    Article  PubMed  CAS  Google Scholar 

  • Ikeda, K.; Takahashi, Y.; Katagiri, S. Effect of medium change on the development of in vitro matured and fertilized bovine oocytes cultured in medium containing amino acids. J. Vet. Med. Sci. 62:121–123; 2000.

    Article  PubMed  CAS  Google Scholar 

  • Jang, G.; Lee, B. C.; Kang, S. K.; Hwang, W. S. Effect of glycosaminoglycans on the preimplantation development of embryos derived from in vitro fertilization and somatic cell nuclear transfer. Reprod. Fertil. Dev. 30:179–185; 2003.

    Article  CAS  Google Scholar 

  • Khurana, N.; Niemann, H. Effects of cryopreservation on glucose metabolism and survival of bovine morulae and blastocysts derived in vitro or in vivo. Theriogenology 54:313–326; 2000a.

    Article  PubMed  CAS  Google Scholar 

  • Khurana, N.; Niemann, H. Energy metabolism in pre-implantation bovine embryos derived in vitro or in vivo. Biol. Reprod. 62:847–856; 2000b.

    Article  PubMed  CAS  Google Scholar 

  • Kim, H.; Lee, G.; Hyun, S., et al. Embryotropic effect of glycosaminoglycans and receptors in development of porcine pre-implantation embryos. Theriogenology 63:1167–1180; 2005.

    Article  PubMed  CAS  Google Scholar 

  • Krisher, R. L.; Lane, M.; Bavister, B. D. Developmental competence and metabolism of bovine embryos cultured in semi-defined and defined culture media. Biol. Reprod. 60:1345–1352; 1999.

    Article  PubMed  CAS  Google Scholar 

  • Kuwayama, M.; Fujikawa, S.; Nagai, T. Ultrastructure of IVM-IVF bovine blastocysts vetrified after equilibration in glycerol 1,2-propanediol using 2-step and 16-step procedures. Cryobiology 31:415–422; 1994.

    Article  PubMed  CAS  Google Scholar 

  • Lane, M. Maybach, J. M.; Hooper, K.; Halser, J. F.; Gardner, D. K. Cryosurvival and development of bovine blastocysts are enhanced by culture with recombinant albumin and hyaluronan. Mol. Reprod. Dev. 64:70–78; 2003.

    Article  PubMed  Google Scholar 

  • Laurent, T. C. Biochemistry of hyaluronan. Acta Otolaryngol. Suppl. 442:7–24; 1987.

    Google Scholar 

  • Lazzari, G.; Wrenzycki, C.; Herrmann, D.; Duchi, R.; Kruip, T.; Niemann, H.; Galli, C. Cellular and molecular deviations in bovine in vitro produced embryos are related to the large offspring syndrome. Biol. Reprod. 67:767–775; 2002.

    Article  PubMed  CAS  Google Scholar 

  • Lee, C. N.; Ax, R. L. Concentration and composition of glycosaminoglycans in the female bovine reproductive tract. J. Dairy Sci. 67:2006–2009; 1984.

    PubMed  CAS  Google Scholar 

  • Lonergan, P.; Rizos, D.; Gutierrez-Adan, A.; Moreira, P. M.; Pintado, B.; De La Fuente, J.; Boland, M. P. Temporal divergence in the pattern of messenger RNA expression in bovine embryos cultured from the zygote to blastocyst stage in vitro or in vivo. Biol. Reprod. 69:1424–1431; 2003.

    Article  PubMed  CAS  Google Scholar 

  • Massip, A.; Mermillod, P.; Wils, C.; Dessy, F. Effects of dilution procedure and culture conditions after thawing on survival of frozen bovine blastocysts produced in vitro. J. Reprod. Fertil. 97:65–69; 1993.

    Article  PubMed  CAS  Google Scholar 

  • Nedambale, T. L.; Dinnyes, A.; Groen, W.; Dobrinsky, J. R.; Tian, X. C.; Yang, X. Comparison on in vitro fertilized bovine embryos cultured in KSOM or SOF and cryopreserved by slow freezing or vitrification. Theriogenology 62:437–449; 2004.

    Article  PubMed  CAS  Google Scholar 

  • Parrish, J. J.; Susko-Parrish, J. L.; Crister, E. S.; Eyestone, W. H.; First, N. L. Bovine in vitro fertilization with frozen-thawed semen. Theriogenology 25:591–600; 1986.

    Article  PubMed  CAS  Google Scholar 

  • Pollard, J. W.; Leibo, S. P. Comparative cryobiology of in vitro and in vivo derived bovine embryos. Theriogenology 39:287 (abstract); 1993.

    Article  Google Scholar 

  • Prather, R. S.; First, N. L. Effect of cytochalasin B and demecolcine on freeze-thaw survival of murine embryos in vitro. J. Exp. Zool. 239:37–40; 1986.

    Article  PubMed  CAS  Google Scholar 

  • Rizos, D.; Fair, T.; Papadopoulos, S.; Boland, M. P.; Lonergan, P. Developmental, qualitative, and ultrastructural differences between ovine and bovine embryos produced in vivo or in vitro. Mol. Reprod. Dev. 62:320–327; 2002.

    Article  PubMed  CAS  Google Scholar 

  • Roth, Z.; Hansen, P. J. Disruption of nuclear maturation and rearrangement of cytoskeletal elements in bovine oocytes exposed to heat shock during maturation. Reproduction 129:235–244; 2005.

    Article  PubMed  CAS  Google Scholar 

  • Ruoslahti, E.; Yamaguchi, Y. Proteoglycans as modulators of growth factor activities. Cell 64:867–869; 1991.

    Article  PubMed  CAS  Google Scholar 

  • Shiedlin, A.; Bigelow, R.; Christopher, W., et al. Evaluation of hyaluronan from different sources: Streptococcus zooepidemicus, rooster comb, bovine vitreous, and human umbilical cord. Biomacromolecules 5:2122–2127; 2004.

    Article  PubMed  CAS  Google Scholar 

  • Stojkovic, M.; Kolle, S.; Peinl, S., et al. Effects of high concentrations of hyaluronan in culture medium on development and survival rates of fresh and frozen-thawed bovine embryos produced in vitro. Reproduction 124:141–153; 2002.

    Article  PubMed  CAS  Google Scholar 

  • Tominaga, K.; Shimizu, M.; Ooyama, S.; Izaike, Y. Effect of lipid polarization by centrifugation at different developmental stages on post-thaw survival of bovine in vitro produced 16-cell embryos. Theriogenology 53:1669–1680; 2000.

    Article  PubMed  CAS  Google Scholar 

  • Toole, B. P.; Zoltan-Jones, A.; Misra, S.; Ghatak, S. Hyaluronan: a critical component of epithelial-mesenchymal and epithelial-carcinoma transitions. Cells Tiss. Organs 179:66–72; 2005.

    Article  CAS  Google Scholar 

  • Vajta, G. Vitrification of the oocytes and embryos of domestic animals. Anim. Reprod. Sci. 60–61:357–364; 2000.

    Article  PubMed  Google Scholar 

  • Valcarcel, A.; de Matos, D. G.; Furnus, C. C. The hyaluronic acid receptor (CD44) is expressed in bovine oocytes and preimplantation stage embryos. Theriogenology 51:193 (abstract); 1999.

    Article  Google Scholar 

  • Yasuda, M.; Nakano, K.; Yasumoto, K.; Tanaka, Y. CD44: functional relevance to inflammation and malignancy. Histol. Histopathol. 17:945–50; 2002.

    PubMed  CAS  Google Scholar 

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  1. Department of Animal Sciences, University of Florida, P.O. Box 110910, 32611-0910, Gainesville, Florida

    M. Franco & P. J. Hansen

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  1. M. Franco
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  2. P. J. Hansen
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Franco, M., Hansen, P.J. Effects of hyaluronic acid in culture and cytochalasin B treatment before freezing on survival of cryopreserved bovine embryos produced in vitro. In Vitro Cell.Dev.Biol.-Animal 42, 40–44 (2006). https://doi.org/10.1007/s11626-006-0010-1

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  • DOI: https://doi.org/10.1007/s11626-006-0010-1

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