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Ultrastructure of bovine in vitro-produced blastocysts cryopreserved by vitrification

Published online by Cambridge University Press:  15 January 2010

Shigeki Ohboshi ,
Noboru Fujihara ,
Tatsuyuki Yoshida  and
Hiroshi Tomagane
Shigeki Ohboshi*
Affiliation:
Nippon Veterinary and Animal Science University, Tokyo, and Kyushu University, Fukuoka, Japan
Noboru Fujihara
Affiliation:
Nippon Veterinary and Animal Science University, Tokyo, and Kyushu University, Fukuoka, Japan
Tatsuyuki Yoshida
Affiliation:
Nippon Veterinary and Animal Science University, Tokyo, and Kyushu University, Fukuoka, Japan
Hiroshi Tomagane
Affiliation:
Nippon Veterinary and Animal Science University, Tokyo, and Kyushu University, Fukuoka, Japan
*
Shigeki Ohboshi, Department of Animal Science, Nippon Veterinary and Animal Science University, 1–7–1 Kyonan-cho, Musashino-shi, Tokyo 180, Japan. Telephone: +81-422-31-4151 (ext. 296). Fax: +81-422-33-2094. e-mail: jag07506@niftyserve.or.jp.
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Summary

The objective of this study was to examine ultrastructural aspects of bovine in vitro-produced blastocysts associated with cryopreservation by vitrification. Morphologically good embryos were used and treated with ethylene-glycol-based vitrification solution (VS). The untreated embryos had conventional fine structure. The post-warming embryos treated with direct exposure to VS (one-step procedure) showed cellular damage structurally by cryopreservation, which included loss of microvilli, disruption of the plasma membrane, mitochondrial changes and swelling of the endoplasmic reticulum. However, nuclei and junctional regions seemed to be resistant to cryoinjury. In contrast, the post-warming embryos pre-equilibra ted with 10% ethylene glycol for 5 min and subsequent exposure to VS (two-step procedure) showed less damage than those treated by the one-step procedure. Post-warming embryos treated by the two-step procedure were cultured in vitro for 18 h. Some embryos survived and their structures re-formed to the former state, while other embryos showed serious injuries and could not reconstitute the blastocoele. Three post-warming embryos treated by the two-step procedure that survived after in vitro culture were transferred to three recipients and one of these resulted in pregnancy. These results indicate that cryopreservation by vitrification can damage membranous structures of the cells of bovine embryos, the extent and nature of this damage being dependent on the vitrification procedure.

Keywords

BlastocystBovineIVFUltrastructureVitrification
Type
Research Article
Information
Zygote , Volume 6 , Issue 1 , February 1998 , pp. 17 - 26
Copyright
Copyright © Cambridge University Press 1998

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References

Ali, J. & Shelton, N. (1993). Successful vitrification of day-6 sheep embryos, J. Reprod. Fertil. 99, 65–70.CrossRefGoogle ScholarPubMed
Douchi, O., Takakura, H. & Imái, K. (1990). Transfer of bovine embryos cryopreserved by vitrification. Jpn. j. Anim. Reprod. 36, 69–72.CrossRefGoogle Scholar
Hemandez-Ledezma, J.J., Mathialagan, N., Villanueva, C., Sikes, J.D. & Roberts, R.M. (1993). Expression of bovine trophoblast interferons by in vitro-derived blastocysts is correlated with their morphological quality and stage of development. Mol. Reprod. Dev. 36, 1–6.CrossRefGoogle Scholar
Heyman, Y., Chesné, P., Chupin, D. & Ménézo, Y. (1987). Improvement of survival rate of frozen cattle blastocysts after transfer with trophoblastic vesicles. Theriogenology 27, 477–84.CrossRefGoogle ScholarPubMed
Iwasaki, S., Yoshikane, Y., Li, X., Watanabe, S. & Nakahara, T. (1994). Effects of freezing of bovine preimplantation embryos derived from oocytes fertilized in vitro on survival of their inner cell mass cells. Mol. Repród. Dev. 37, 272–5.CrossRefGoogle ScholarPubMed
Kasai, M., Komi, J.H., Takakamo, A., Tsudera, H., Sakurai, T. & Machida, T. (1990). A simple method for mouse embryo cryopreservation in a low toxicity vitrification solution, without appreciable loss of viability, J. Reprod. Fertil. 89, 91–7.CrossRefGoogle Scholar
Kuwayama, M., Hamáno, S. & Nagai, T. (1992). Vitrification of bovine blastocysts obtained by in vitro culture of oocytes matured and fertilized in vitro. J. Reprod. Fertil. 96, 187–93.CrossRefGoogle ScholarPubMed
Kuwayama, M., Fujikawa, S. & Nagai, T. (1994). Ultrastructure of IVM-IVF bovine blastocysts vitrified after equilibration in glycerol 1, 2-propanediol using 2-step and 16-step procedures. Cryobiology 31, 415–22.CrossRefGoogle ScholarPubMed
Linares, T. & Plöen, L. (1981). On the ultrastructure of seven day old normal (blastocysts) and normal bovine embryos. Zbl. Vet. Med. C. Ant. Histol. Embryol. 10, 212–26.CrossRefGoogle ScholarPubMed
Mahmoudzadeh, A.R., Van Soom, A., Bols, P., Ysebaert, M.T. & de Kruif, A. (1995). Optimization of a simple vitrification procedure for bovine embryos produced in vitro: effect of developmental stage, two-step addition of cryoprotectant and sucrose dilution on embryonic survival. j. Reprod. Fertil. 103, 33–9.CrossRefGoogle ScholarPubMed
Massip, A., Van der Zwalmen, P., Scheffen, B. & Ectors, F. (1986). Pregnancies following transfer of cattle embryos preserved by vitrification. Cryo-Lett. 7, 270–3.Google Scholar
Massip, A., Van der Zwalmen, P., Scheffen, B. & Ectors, F. (1989). Some significant steps in the cryopreservation of mammalian embryos with a note on a vitrification procedure. Anim. Repród. Sci. 19, 117–29.CrossRefGoogle Scholar
Massip, A., Mermillod, P., Wils, C. & Dessey, F. (1993). Effects of dilution procedure and culture conditions after thawing on survival of frozen bovine blasotcysts produced in vitro. J. Repród. Fértil. 97, 65–9.CrossRefGoogle Scholar
Massip, A., Mermillod, P. & Dinnyés, A. (1995). Morphology and biochemistry of in vitro-produced bovine embryos: implications for their cryopreservation. Hum. Repród. 10, 3004–11.CrossRefGoogle ScholarPubMed
Mohr, L.R. & Trounson, A.O. (1981). Structural changes associated with freezing of bovine embryos. Biol. Repród. 25, 1009–25.CrossRefGoogle ScholarPubMed
Ohboshi, S., Nakamichi, R., Hanada, K., Zhao, J., Hattori, M., Fujihara, N. & Umetsu, R. (1995). Ultrastructural comparison of bovine blastocysts developed in vivo and in vitro. Asian-Austral. J. Anim. Sci. 8, 599–605.Google Scholar
Ohboshi, S., Eton, T., Sakamoto, K., Fujihara, N., Yoshida, T. & Tomogane, H. (1997a). Effects of bovine serum proteins in culture medium on post-warming survival of bovine blastocysts developed in vitro. Theriogenology 47, 1237–43.CrossRefGoogle ScholarPubMed
Ohboshi, S., Fujihara, N., Yoshida, T. & Tomogane, H. (1997b). Usefulness of polyethylene glycol for cryopreservation by vitrification of in vitro-derived bovine blastocysts. Anim. Reprod. Sci. 48, 27–36.CrossRefGoogle ScholarPubMed
Rall, W.F. (1987). Factors affecting the survival of mouse embryos cryopreserved by vitrification. Cryobiology 24, 387–402.CrossRefGoogle ScholarPubMed
Rall, W.F. & Fahy, G.M. (1985). Ice-free cryopreservation of mouse embryos at –196°C by vitrification. Nature 313, 573–5.CrossRefGoogle Scholar
Rieger, D., Bruyas, J.-F., Lagneaux, D., Bézard, J. & Palmer, E. (1991). The effects of cryopreservation on the metabolic activity of day-6.5 horse embryos, J. Reprod. Fertil. Suppl. 44, 411–17.Google ScholarPubMed
Rieger, D., Pollard, J.W. & Leibo, S.P. (1993). The effect of cryopreservation on the metabolic activity of in vitro produced cattle blastocysts. Cryobiology 30, 631.Google Scholar
Rieger, D., Grisait, B., Semple, E., Van Langendonckt, A., Betteridge, K.J. & Dessy, F. (1995). Comparison of the effects of oviductal cell co-culture and oviductal cell-conditioned medium on the development and metabolic activity of cattle embryos, J. Reprod. Fertil. 105, 91–8.CrossRefGoogle ScholarPubMed
Rubinsky, B., Arav, A. & Devries, A.L. (1992). The cryo-protective effect of antifreeze glycopeptides from antarctic fishes. Cryobiology 29, 69–77.CrossRefGoogle Scholar
Scheffen, B., Van Der Zwalmen, P. & Massip, A. (1986). A simple and efficient procedure for preservation of mouse embryos by vitrification. Cryo-Lett. 7, 260–9.Google Scholar
Tachikawa, S., Otoi, T., Kondo, S., Machida, T. & Kasai, M. (1993). Successful vitrification of bovine blastocysts, derived by in vitro maturation and fertilization. Mol. Repród. Dev. 34, 266–71.CrossRefGoogle ScholarPubMed
Takahashi, Y. & First, N.L. (1992). In vitro development of bovine one-cell embryos: influence of glucose, lactate, pyruvate, amino acids and vitamins. Theriogenology 37, 963–78.CrossRefGoogle ScholarPubMed
Valdes, C.A., Abas Mazni, O., Kanagawa, H. & Fujikawa, S. (1990). Confirmation by cryoelectron microscopy of the absence of crystallization using a vitrification solution. Cryo-Lett. 11, 351–8.Google Scholar
Wilson, J.M., Caceci, T., Potter, G.D. & Kraemer, D.C. (1987). Ultrastructure of cryopreserved horse embryos, j. Reprod. Fertil. Suppl. 35, 405–17.Google Scholar
Wurth, Y.A., Reinders, J.M.C., Rall, W.F. & Kruip, Th.A.M. (1994). Developmental potential of in vitro produced bovine embryos following cryopreservation and single-embryo transfer. Theriogenology 42, 1275–84.CrossRefGoogle Scholar