Hostname: page-component-cd9895bd7-jkksz Total loading time: 0 Render date: 2024-12-29T06:30:08.194Z Has data issue: false hasContentIssue false

Effect of vitrification technique and assisted hatching on rabbit embryo developmental rate

Published online by Cambridge University Press:  01 February 2009

M. Popelková
Affiliation:
Centre of Assisted Reproduction, IInd Gynaecological-Obstetrical Clinic of the L. Pasteur, Faculty Hospital and Medical Faculty of the P.J. Šafárik University, Rastislavova 43, 041 90 Košice, Slovak Republic.
Z. Turanová
Affiliation:
Department of Animal Genetics and Reproduction, Research Institute of Animal Production, Slovak Agricultural Research Centre, Hlohovská 2, 949 92 Nitra, Slovak Republic.
L. Koprdová
Affiliation:
Department of Animal Genetics and Reproduction, Research Institute of Animal Production, Slovak Agricultural Research Centre, Hlohovská 2, 949 92 Nitra, Slovak Republic. Department of Zoology and Anthropology, Faculty of Nature Sciences, Constantine the Philosopher University, Nitra, Slovak Republic.
A. Ostró
Affiliation:
Centre of Assisted Reproduction, IInd Gynaecological-Obstetrical Clinic of the L. Pasteur, Faculty Hospital and Medical Faculty of the P.J. Šafárik University, Rastislavova 43, 041 90 Košice, Slovak Republic.
S. Toporcerová
Affiliation:
Centre of Assisted Reproduction, IInd Gynaecological-Obstetrical Clinic of the L. Pasteur, Faculty Hospital and Medical Faculty of the P.J. Šafárik University, Rastislavova 43, 041 90 Košice, Slovak Republic.
A. V. Makarevich
Affiliation:
Department of Animal Genetics and Reproduction, Research Institute of Animal Production, Slovak Agricultural Research Centre, Hlohovská 2, 949 92 Nitra, Slovak Republic.
P. Chrenek*
Affiliation:
RIAP SARC, Hlohovská 2, 949 92 Nitra, Slovak Republic. Centre of Assisted Reproduction, IInd Gynaecological-Obstetrical Clinic of the L. Pasteur, Faculty Hospital and Medical Faculty of the P.J. Šafárik University, Rastislavova 43, 041 90 Košice, Slovak Republic. Department of Animal Genetics and Reproduction, Research Institute of Animal Production, Slovak Agricultural Research Centre, Hlohovská 2, 949 92 Nitra, Slovak Republic.
*
All correspondence to: Peter Chrenek. RIAP SARC, Hlohovská 2, 949 92 Nitra, Slovak Republic. Tel: +421 37 6546 236. Fax: +421 37 6546 189. e-mail: chrenekp@yahoo.com

Summary

The aim of the study was to determine the efficiency of two vitrification techniques followed by two assisted hatching (AH) techniques based on post-thaw developmental capacity of precompacted rabbit embryos and their ability to leave the zona pellucida (hatching) during in vitro culture. The total cell number and embryo diameter as additional markers of embryo quality after warming were evaluated. In vivo fertilized, in vitro cultured 8–12-cell rabbit embryos obtained from superovulated rabbit does were cryopreserved by two-step vitrification method using ethylene glycol (EG) as cryoprotectant or by one-step vitrification method with EG and Ficoll (EG+Ficoll). Thawed embryos were subjected to enzymatic or mechanical AH. Vitrified EG group showed significantly lower (P < 0.05) blastocyst rate (22.5%) and hatching rate (15%) than those vitrified with EG + Ficoll (63 and 63% resp.) and that of control (97 and 97% respectively). Significantly lower values of total cell number (P < 0.05) as well as embryo diameter (P < 0.01) in EG group compared with EG + Ficoll and control group were recorded. No significant difference was found in developmental potential of warmed embryos treated by either mechanical or enzymatic AH. The present study demonstrates that the EG + Ficoll vitrification protocol provides superior embryo survival rates over the EG vitrification protocol for 8–12-cell stage precompacted rabbit embryos. No positive effect of either mechanical or enzymatic AH on the post-thaw viability and quality of rabbit embryos in vitro was observed.

Type
Research Article
Copyright
Copyright © Cambridge University Press 2008

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Balaban, B., Urman, B., Alatas, C., Mercan, R., Mumcu, A. & Isiklar, A. (2002). A comparison of four different techniques of assisted hatching. Hum. Reprod. 17, 1239–43.CrossRefGoogle ScholarPubMed
Blake, D.A., Forsberg, A.S., Johansson, B.R. & Wikland, M. (2001). Laser zona pellucida thinning – an alternative approach to assisted hatching. Hum. Reprod. 16, 1959–64.CrossRefGoogle ScholarPubMed
Chrenek, P. & Makarevich, A. (2005). Production of rabbit chimeric embryos by aggregation of zona-free nuclear transfer blastomeres. Zygote 13, 3944.CrossRefGoogle ScholarPubMed
De Vos, A. & Van Steirteghem, A. (2000). Zona hardening, zona drilling and assisted hatching: new achievements in assisted reproduction. Cells Tissues Organs 166, 220–7.CrossRefGoogle ScholarPubMed
Fabian, D., Gjorret, J.O., Berthelot, F., Martinat-Botté, F. & Maddox-Hytel, P. (2005). Ultrastructure and cell death of in vivo derived and vitrified porcine blastocysts. Mol. Reprod. Dev. 70 (2), 155–65.CrossRefGoogle ScholarPubMed
Hredzák, R., Ostró, A., Ždilová, V., Maraček, I. & Kačmárik, J. (2006). Survival of mouse embryos after vitrification depending on the cooling rate of the cryoprotectant solution. Acta Veterinaria Hungarica 54 (1), 117–25.CrossRefGoogle ScholarPubMed
Kaidi, S., Bernard, S., Lambert, P., Massip, A., Dessy, F. & Donnay, I. (2001). Effect of conventional controlled-rate freezing and vitrification on morphology and metabolism of bovine blastocysts produced in vitro. Biol. Reprod. 65, 1127–34.CrossRefGoogle ScholarPubMed
Kong, I.K., Lee, S.I., Im, Y.T., Cho, S.G., Ohh, H.J., Ohh, D.H. & Bae, I.H. (2000). Improvement of post-thaw hatching rates of in vitro-produced bovine embryos vitrified by ultra-mini straw (abstract). Theriogenology 53, 258.Google Scholar
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.CrossRefGoogle ScholarPubMed
Kuleshova, L.L. & Lopata, A. (2002). Vitrification can be more favourable than slow cooling. Fertil. Steril. 78, 449–54.CrossRefGoogle ScholarPubMed
Liebermann, J., Dietl, J. & Vanderzwalmen, P. (2003). Recent developments in human oocyte, embryo and blastocyst vitrification: where are we now? Reprod. Biomed. Online 7, 623–33.CrossRefGoogle ScholarPubMed
Makarevich, A.V., Chrenek, P. & Fl'ak, P. (2006). The influence of microinjection of foreign gene into the pronucleus of fertilized egg on the preimplantation development, cell number and diameter of rabbit embryos. Asian-Aust. J. Anim. Sci. 19 (2), 171–5.CrossRefGoogle Scholar
Mori, M., Otoi, T. & Suzuki, T. (2002). Correlation between the cell number and diameter in bovine embryos produced in vitro. Reprod. Dom. Anim. 37, 181–4.CrossRefGoogle ScholarPubMed
Naik, B.R., Rao, B.S., Vagdevi, R., Gnanprakash, M., Amarnath, D. & Rao, V.H. (2005). Conventional slow freezing, vitrification and open pulled straw (OPS) vitrification of rabbit embryos. Anim. Reprod. Sci. 86, 329–38.CrossRefGoogle ScholarPubMed
Ng, E.H.Y., Naveed, F., Lau, E.Y.L., Yeung, W.S.B., Chan, C.Ch.W., Tang, O.S. & Ho, P.Ch. (2005). A randomized double-blind controlled study of the efficacy of laser-assisted hatching on implantation and pregnancy rates of frozen–thawed embryo transfer at the cleavage stage. Hum. Reprod. 20 (4), 979–85.CrossRefGoogle ScholarPubMed
Papis, K., Sypecka, J., Korwin-Kossakowski, M., Wenta-Muchalska, E., Bilska, B. (2005). Banking of embryos of mutated, paralytic tremor rabbit by means of vitrification. Laboratory Animals 39, 284–9.CrossRefGoogle ScholarPubMed
Popelková, M., Chrenek, P., Pivko, J., Makarevich, A.V., Kubovičová, E. & Kačmárik, J. (2005). Survival and ultrastructure of gene-microinjected rabbit embryos after vitrification. Zygote 13, 283–93.CrossRefGoogle ScholarPubMed
Smorag, Z., Gajda, B., Wieczorek, B. & Jura, J. (1989). Stage dependent viability of vitrified rabbit embryos. Theriogenology 31, 1227–31.CrossRefGoogle ScholarPubMed
Stehlik, E., Stehlik, J., Katayama, K.P., Kuwayama, M., Jambor, V., Brohammer, R. & Kato, O. (2005). Vitrification demonstrates significant improvement versus slow freezing of human blastocysts. Reprod. Biomed. Online 11, 53–7.CrossRefGoogle ScholarPubMed
Vanderzwalmen, P., Bertin, G., Debauche, Ch., Standaert, V., Bollen, N., Van Roosendaal, E., Vandervorst, M., Schoysman, R. & Zech, N. (2003). Vitrification of human blastocysts with Hemi-Straw carrier: application of assisted hatching after thawing. Hum. Reprod. 18, 1504–11.CrossRefGoogle ScholarPubMed
Vicente, J.S. & Garcia-Ximénez, F. (1994). Osmotic and cryoprotective effects of a mixture of DMSO and ethylene glycol on rabbit morulae. Theriogenology 42, 1205–15.CrossRefGoogle ScholarPubMed
Vicente, J.S., Viudes-De-Castro, M.P., Garcia, M.L. & Baselga, M. (2003). Effect of rabbit line on a program of cryopreserved embryos by vitrification. Reprod. Nutr. Dev. 43, 137–43.CrossRefGoogle ScholarPubMed
Zhao, X.M., Quan, G.B., Zhou, G.B., Hou, Y.P. & Zhu, S.E. (2007). Conventional freezing, straw, and open-pulled straw vitrification of mouse two pronuclear (2-PN) embryos. Anim. Biotechnol. 18, 203–12.CrossRefGoogle Scholar
Zhou, G.B., Hou, Y.P., Jin, F., Yang, Q.E., Yang, Z.Q., Quan, G.B., Tan, H.M. & Zhu, S.E. (2005). Vitrification of mouse embryos at various stages by open-pulled straw (OPS) method. Anim. Biotechnol. 16, 153–63.CrossRefGoogle ScholarPubMed
Zhou, C., Zhou, G.B., Zhu, S.E., Hou, Y.P., Jin, F., Zhao, X.M. & Hong, Q.H. (2007). Open-pulled (OPS) vitrification of mouse hatched blastocysts. Anim. Biotechnol. 18, 4554.CrossRefGoogle ScholarPubMed