Skip to main content

Advertisement

Log in

Animal oocyte and embryo cryopreservation

  • Review Paper
  • Published:
Cell and Tissue Banking Aims and scope Submit manuscript

Abstract

Cryopreservation of oocytes and embryos is a crucial step for the widespread and conservation of animal genetic resources. However, oocytes and early embryos are very sensitive to chilling and cryopreservation and although new advances have been achieved in the past few years the perfect protocol has not yet been established. All oocytes and embryos suffer considerable morphological and functional damage during cryopreservation but the extent of the injury as well as differences in survival and developmental rates may be highly variable depending on the species, developmental stage and origin (for example, in vitro produced or in vivo derived, micromanipulated or not). Currently, there are two methods for gamete and embryos cryopreservation: slow freezing and vitrification. We have experienced both techniques but vitrification has become a viable and promising alternative to traditional approaches especially when dealing with in vitro produced or micromanipulated embryos and oocytes. Recently new strategies based on emerging studies in the field of lipid research have been used to reduce intracellular lipid content in bovine in vitro produced embryos and therefore increase their tolerance to micromanipulation and cryopreservation. The addition of a conjugated isomer of linoleic acid, the trans-10, cis-12 octadecadienoic acid to embryo culture medium more than twice improved embryo post-thawing viability after micromanipulation and vitrification. Vitrification was also used for the cryopreservation of embryos belonging to the Portuguese Animal Germplasm Bank project presently running at our facilities.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

Similar content being viewed by others

References

  • Abe H, Matsuzaki S, Hoshi H (2002a) Ultrastructural differences in bovine morulae classified as high and low qualities by morphological evaluation. Theriogenology 57:1273–1283

    Article  PubMed  CAS  Google Scholar 

  • Abe H, Yamashita S, Itoh T, Satoh T, Hoshi H (2002b) Accumulation of cytoplasmatic lipid droplets in bovine embryos and cryotolerance of embryos developed in different culture systems using serum-free medium or in serum-containing medium. Mol Reprod Dev 61:57–66

    Article  PubMed  CAS  Google Scholar 

  • Andrabi SMH, Maxwell WMC (2007) A review on reproductive biotechnologies for conservation of endangered mammalian species. Anim Reprod Sci 99:223–243

    Article  PubMed  CAS  Google Scholar 

  • Arav A, Yavin S, Zeron Y, Natan D, Dekel I, Gacitua H (2002) New trends in gamete`s cryopreservation. Mol Cell Endocrinol 187:77–81

    Article  PubMed  CAS  Google Scholar 

  • Berthelot F, Marinat-Botte F, Locatelli A, Perreau C, Terqui M (2000) Piglets born after vitrification of embryos using the open pulled straw method. Cryobiology 41:116–124

    Article  PubMed  CAS  Google Scholar 

  • Carvalhais I, Marques CC, Baptista MC, Vasques MI, Horta AEM, Pereira RM (2006) Effect of trans-10, cis-12 conjugated linoleic acid (CLA) on post-thaw viability of biopsied bovine embryos. Reprod Dom Anim 41:334–335

    Google Scholar 

  • Checura CM, Seidel GE (2007) Effect of macromolecules in solutions for vitrification of mature bovine oocytes. Theriogenology 67:919–930

    Article  PubMed  CAS  Google Scholar 

  • Cho SR, Cho SK, Lee SL, Choe SY, Rho GJ (2002) Enhanced cryosurvival of bovine blastocysts produced in vitro in serum-free medium. J Assist Reprod Genet 19:487–492

    Article  PubMed  Google Scholar 

  • Cuello C, Berthelot F, Delaleu B, Venturi E, Pastor LM, Vazquez JM, Roca J, Martinat-Botte F, Martinez EA (2007) The effectiveness of the stereomicroscopic evaluation of embryo quality in vitrified–warmed porcine blastocysts: an ultrastructural and cell death study. Theriogenology 67:970–982

    Article  PubMed  CAS  Google Scholar 

  • De Matos DG, Gasparrini B, Pasqualini SR, Thompson JG (2002) Effect of glutathione synthesis stimulation during in vitro maturation of ovine oocytes on embryo development and intracellular peroxide content. Theriogenology 57:1443–1451

    Article  PubMed  Google Scholar 

  • Diez C, Heyman Y, Bourhis D, Guyader-Joly C, Degrouard J, Renard JP (2001) Delipidating in vitro-produced bovine zygotes: effect on further development and consequences for freezability. Theriogenology 55:923–936

    Article  PubMed  CAS  Google Scholar 

  • Eckert J, Niemann H (1995) In vitro maturation, fertilization and culture to blastocysts of bovine oocytes in protein-free media. Theriogenology 43:1211–1225

    Article  PubMed  CAS  Google Scholar 

  • Eroglu A, Lawitts JA, Toner M, Toth TL (2003) Quantitative microinjection of trehalose into mouse oocytes and zygotes, and its effect on development. Cryobiology 46:121–134

    Article  PubMed  CAS  Google Scholar 

  • Fair T, Lonergan P, Dinnyes A, Cottell DC, Hyttel P, Ward FA, Boland MP (2001) Ultrastructure of bovine blastocysts following cryopreservation: effect of method of blastocyst production. Mol Reprod Dev 58:186–195

    Article  PubMed  CAS  Google Scholar 

  • Feugang JM, de Roover R, Moens A, Léonard S, Dessy F, Donnay I (2004) Addition of beta-mercaptoethanol or Trolox at the morula/blastocyst stage improves the quality of bovine blastocysts and prevents induction of apoptosis and degeneration by prooxidant agents. Theriogenology 61:71–90

    Article  PubMed  CAS  Google Scholar 

  • Fuchinoue K, Fukunaga N, Chiba S, Nakajo Y, Yagi A, Kyono K (2004) Freezing of human immature oocytes using cryoloops with Taxol in the vitrification solution. J Assist Reprod Genet 21:307–309

    Article  PubMed  Google Scholar 

  • Fujihira T, Kishida R, Fukui Y (2004) Developmental capacity of vitrified immature porcine oocytes following ICSI: effects of cytochalasin B and cryoprotectants. Cryobiology 49:286–290

    Article  PubMed  CAS  Google Scholar 

  • Gama LT (2006) Animal genetic resources and sustainable development in the Mediterranean region. In: Ramalho Ribeiro JM, Horta AEM, Mosconi C, Rosati A (eds) Animal products from the Mediterranean area. EAAP pub. 119. Wageningen Academic Publishers, pp 127–135

  • Genicot G, Leroy JL, Van Soom A, Donnay I (2005) The use of a fluorescent dye, Nile red, to evaluate the lipid content of single mammalian oocytes. Theriogenology 63:1181–1194

    Article  PubMed  CAS  Google Scholar 

  • Harvey MB, Arcellana-Panlilio MY, Zhang X, Schultz GA, Watson AJ (1995) Expression of genes encoding antioxidant enzymes in preimplantation mouse and cow embryos and primary bovine oviduct cultures employed for embryo coculture. Biol Reprod 53:532–540

    Article  PubMed  CAS  Google Scholar 

  • Hasler JF, Henderson WB, Hurtgen PJ, Jin ZQ, McCauley AD, Mower SA, Neely B, Shuey LS, Stokes JE, Trimmer SA (1995) Production, freezing and transfer of bovine IVF embryos and subsequent calving results. Theriogenology 43:141–152

    Article  Google Scholar 

  • Hochi S, Fujimoto T, Oguri N (1995) Large equine blastocysts are damaged by vitrification procedures. Reprod Fertil Dev 7:113–117

    Article  PubMed  CAS  Google Scholar 

  • Hochi S, Kimura K, Hanada A (1999) Effect of linoleic acid-albumin in the culture medium on freezing sensitivity of in vitro-produced bovine morulae. Theriogenology 52:497–504

    Article  PubMed  CAS  Google Scholar 

  • Horta AEM (2007) Banco Português de Germoplasma Animal. In: 6th Relatório de Progresso do Projecto AGRO438/2003090023163, pp 28

  • Isachenko V, Isachenko E, Michelmann HW, Alabart JL, Vazquez I, Bezugly N, Nawroth F (2001) Lipolysis and ultrastructural changes of intracellular lipid vesicles after cooling of bovine and porcine GV-oocytes. Anat Histol Embryol 30:333–338

    Article  PubMed  CAS  Google Scholar 

  • Jain JK, Paulson RJ (2006) Oocyte cryopreservation. Fertil Steril 86:1037–1046

    Article  PubMed  CAS  Google Scholar 

  • Kuwayama M, Vajta G, Ieda S, Kato O (2005) Comparison of open and closed methods for vitrification of human embryos and the elimination of potential contamination. Reprod Biomed Online 11:608–614

    PubMed  Google Scholar 

  • Lane M, Bavister BD, Lyons EA, Forest KT (1999) Containerless vitrification of mammalian oocytes and embryos. Nat Biotechnol 17:1234–1236

    Article  PubMed  CAS  Google Scholar 

  • Laowtammathron C, Lorthongpanich C, Ketudat-Cairns M, Hochi S, Parnpai R (2005) Factors affecting cryosurvival of nuclear-transferred bovine and swamp buffalo blastocysts: effects of hatching stage, linoleic acid-albumin in IVC medium and Ficoll supplementation to vitrification solution. Theriogenology 64:1185–1196

    Article  PubMed  CAS  Google Scholar 

  • Lapa M, Marques CC, Baptista MC, Vasques MI, Horta AEM, Portugal PV, Bessa RJB, Pereira RM (2005) Fatty acid profile of oocytes and blastocysts during in vitro bovine embryo production. Reprod Dom Anim 40:365

    Google Scholar 

  • Ledda S, Leoni G, Bogliolo L, Naitana S (2001) Oocyte cryopreservation and ovarian tissue banking. Theriogenology 55:1359–1371

    Article  PubMed  CAS  Google Scholar 

  • LeRoy JL, Genicot G, Donnay I, Van Soom A (2005) Evaluation of the lipid content in bovine oocytes and embryos with nile red: a practical approach. Reprod Domest Anim 40:76–78

    Article  PubMed  CAS  Google Scholar 

  • Lim JM, Fukui Y, Ono H (1992) Developmental competence of bovine oocytes at various maturation stages followed by in vitro maturation and fertilization. Theriogenology 37:351–362

    Article  Google Scholar 

  • Marques CC, Baptista MC, Vasques MI, Horta AEM, Pereira RM (2007) Effect of polyunsaturated fatty acids (PUFA) on bovine oocyte in vitro maturation and subsequent embryo development and freezability. Reprod Dom Anim 42:108–109

    Google Scholar 

  • Martínez AG, Valcácel A, Furnus CC, de Matos DG, Iorio G, de las Heras MA (2006) Cryopreservation of in vitro produced bovine embryos. Small Rum Res 63:288–296

    Article  Google Scholar 

  • Martino A, Songsasen N, Leibo SP (1996) Development into blastocysts of bovine oocytes cryopreserved by ultrarapid cooling. Biol Reprod 54:1059–1069

    Article  PubMed  CAS  Google Scholar 

  • Mavrides A, Morrol D (2005) Bypassing the effect of zona pellucida changes on embryo formation following cryopreservation of bovines oocytes. Eur J Obstet Gynecol 18:66–70

    Article  Google Scholar 

  • McEvoy TG, Coull GD, Broadbent PJ, Hutchinson JSM, Speake BK (2000) Fatty acid composition of lipids in immature cattle, pig and sheep oocytes with intact zona pellucida. J Reprod Fertil 118:163–170

    Article  PubMed  CAS  Google Scholar 

  • McEvoy TG, Robinson JJ, Sinclair KD (2001) Developmental consequences of embryo and cell manipulation in mice and farm animals. Reproduction 122:507–518

    Article  PubMed  CAS  Google Scholar 

  • Men H, Agca Y, Riley L, Critser JK (2006) Improved survival of vitrified porcine embryos after partial delipation through chemically stimulated lipolysis and inhibition of apoptosis. Theriogenology 66:2008–2016

    Article  PubMed  CAS  Google Scholar 

  • Mermillod P, Traldi AL, Baril G, Beckers JF, Massip A, Cognie´ Y (1997) A vitrification method for direct transfer of sheep embryos. In: Proceedings of the 15th science meeting on European embryo transfer association, p 212

  • Morato R, Izquierdo D, Albarracin JL, Anguita B, Palomo MJ, Jimenez-Macedo AR, Paramio MT, Mogas T (2008) Effects of pre-treating in vitro-matured bovine oocytes with the cytoskeleton stabilizing agent taxol prior to vitrification. Mol Reprod Dev 75:191–201

    Article  PubMed  CAS  Google Scholar 

  • Nagashima H, Cameron RDA, Kuwama M, Young M, Beebe L, Blackshaw AW, Nottle MB (1999) Survival of porcine delipated oocytes and embryos after cryopreservation by freezing and vitrification. J Reprod Dev 45:167–176

    Article  Google Scholar 

  • Nasr-Esfahani MH, Aitken JR, Johnson MH (1990) Hydrogen peroxide levels in mouse oocytes and early cleavage stage embryos developed in vitro or in vivo. Development 109:501–507

    PubMed  CAS  Google Scholar 

  • Nedambale T, Du F, Yang X, Tian X (2006) Higher survival rate of vitrified and thawed in vitro produced bovine blastocysts following culture in defined medium supplemented with β-mercaptoethanol. Anim Reprod Sci 93:61–75

    Article  PubMed  CAS  Google Scholar 

  • Nieman H, Rath D (2001) Progress in reproductive biotechnology in swine. Theriogenology 56:1291–1304

    Article  Google Scholar 

  • Pangestu M, Lewis I, Shaw J (1996) Alpha tocopherol microinjection protects in vitro-produced bovine pronuclear-stage embryos from chill and cryo-injury. In: Proceedings of the 13th international congress on animal reproduction (ICAR), vol 3, Sydney, pp 15–17

  • Pereira RM, Baptista MC, Vasques MI, Horta AEM, Portugal PV, Bessa RJB, Marques CC (2004) Post-thawing resistance of bovine embryos is improved by trans-10 cis-12 conjugated linoleic acid (CLA). In: Proceedings of 15th international congress on animal reproduction (ICAR), vol 2, Porto Seguro-Brazil, pp 538

  • Pereira RM, Baptista MC, Vasques MI, Horta AEM, Portugal PV, Bessa RJB, Chagas e Silva J, Silva Pereira M, Marques CC (2007) Cryo-survival of bovine blastocysts is enhanced by culture with trans-10 cis-12 conjugated linoleic acid (10t, 12c CLA). Anim Reprod Sci 98:293–301

    Article  PubMed  CAS  Google Scholar 

  • Pereira RM, Carvalhais I, Pimenta J, Baptista MC, Vasques MI, Horta AEM, Santos IC, Marques MR, Reis A, Silva Pereira M, Marques CC (2008) Biopsied and vitrified bovine embryos viability is improved by trans10, cis12 conjugated linoleic acid supplementation during in vitro embryo culture. Anim Reprod Sci 106:322–332

    Article  PubMed  CAS  Google Scholar 

  • Pinyopummintr T, Bavister BD (1991) In vitro-matured/in vitro-fertilized bovine oocytes can develop into morulae/blastocysts in chemically defined, protein-free culture media. Biol Reprod 45:736–42

    Article  PubMed  CAS  Google Scholar 

  • Reis A, Rooke JA, McCallum GJ, Staines ME, Ewen M, Lomax MA, McEvoy TG (2003) Consequences of exposure to serum, with or without vitamin E supplementation, in terms of the fatty acid content and viability of bovine blastocysts produced in vitro. Reprod Fertil Dev 15:275–284

    Article  PubMed  CAS  Google Scholar 

  • Rojas C, Palomo MJ, Albarracın JL, Mogas T (2004) Vitrification of immature and in vitro matured pig oocytes: study of distribution of chromosomes, microtubules, and actin microfilaments. Cryobiology 49:211–220

    Article  PubMed  CAS  Google Scholar 

  • Sata R, Tsujii H, Abe H, Yamashita S, Hochi H (1999) Fatty acid composition of bovine embryo cultured in serum free and serum containing medium during early embryonic development. J Reprod Dev 45:97–103

    Article  CAS  Google Scholar 

  • Seidel GE (2006) Modifying oocytes and embryos to improve their cryopreservation. Theriogenology 65:228–235

    Article  PubMed  CAS  Google Scholar 

  • Shi WQ, Zhu SE, Zhang D, Wang W, Tang G, Hou Y, Tian S (2006) Improved development by Taxol pretreatment after vitrification of in vitro matured porcine oocytes. Reproduction 131:795–804

    Article  PubMed  CAS  Google Scholar 

  • Silvestre MA, Yaniz J, Salvador I, Santolaria P, Lopez-Gatius F (2006) Vitrification of pre-pubertal ovine cumulus–oocyte complexes: effect of cytochalasin B pre-treatment. Anim Reprod Sci 93:176–182

    Article  PubMed  CAS  Google Scholar 

  • Steel R, Hasler JF (2004) Pregnancy rates from transfer of fresh and frozen Holstein and Jersey embryos. Reprod Fertil Dev 16:182–3

    Article  Google Scholar 

  • Succu S, Leoni GG, Berlinguer F, Madeddu M, Bebbere D, Mossa F, Bogliolo L, Ledda S, Naitana S (2007) Effect of vitrification solutions and cooling upon in vitro matured prepubertal ovine oocytes. Theriogenology 68:107–114

    Article  PubMed  CAS  Google Scholar 

  • Taniguchi M, Ikeda A, Arikawa E, Wongsrikeao P, Agung B, Naoi H, Nagai T, Otoi T (2007) Effect of cryoprotectant composition on in vitro viability of in vitro fertilized and cloned bovine embryos following vitrification and in-straw dilution. J Reprod Dev 53:963–969

    Article  PubMed  CAS  Google Scholar 

  • Tominaga K (2004) Cryopreservation and sexing of in vivo- and in vitro-produced bovine embryos for their practical use. J Reprod Dev 50:29–38

    Article  PubMed  Google Scholar 

  • Tominaga K, Shimizu M, Ooyama S, Izaike Y (2000) 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

    Article  PubMed  CAS  Google Scholar 

  • Ushijima H, Yamakawa H, Nagashima H (1999) Cryopreservation of bovine pre-morula-stage in vitro matured/in vitro fertilized embryos after delipidation and before use in nucleus transfer. Biol Reprod 60:534–539

    Article  PubMed  CAS  Google Scholar 

  • Vajta G, Nagy ZP (2006) Are programmable freezers still needed in the embryo laboratory? Review on vitrification. Reprod Biomed Online 12:779–796

    Article  PubMed  Google Scholar 

  • Vajta G, Hyttel P, Callensen H (1997) Morphological changes of in vitro-produced bovine blastocysts after vitrification, in straw direct rehydration, and culture. Mol Reprod Dev 48:9–17

    Article  PubMed  CAS  Google Scholar 

  • Vajta G, Holm P, Kuwayama M, Booth PJ, Jacobsen H, Greve T, Callesen H (1998) Open pulled straw (OPS) vitrification: a new way to reduce cryoinjuries of bovine ova and embryos. Mol Reprod Dev 51:53–58

    Article  PubMed  CAS  Google Scholar 

  • Van Langendonckt A, Morales H, Massip A, Dessy F (1998) Effect of hydrogen peroxide on in vitro development of bovine embryos. Theriogenology 49:221

    Article  Google Scholar 

  • Woelders H, Zuidberg CA, Hiemstra SJ (2006) Animal genetic resources conservation in the Netherlands and Europe: poultry perspectives. Poultry Sci 85:216–222

    CAS  Google Scholar 

  • Zech NH, Lejeune B, Zech H, Vanderzwalmen P (2005) Vitrification of hatching and hatched human blastocysts: effect of an opening in the zona pellucida before vitrification. Reprod Biomed Online 11:355–61

    Article  PubMed  CAS  Google Scholar 

  • Zeron Y, Ocheretny A, Kedar O, Borochov A, Sklan D, Arav A (2001) Seasonal changes in bovine fertility: relation to developmental competence of oocytes, membrane properties and fatty acid composition of follicles. Reproduction 121:447–454

    Article  PubMed  CAS  Google Scholar 

  • Zeron Y, Sklan D, Arav A (2002) Effect of polyunsaturated fatty acid supplementation on biophysical parameters and chilling sensitivity of ewe oocytes. Mol Reprod Dev 61:271–278

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to R. M. Pereira.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Pereira, R.M., Marques, C.C. Animal oocyte and embryo cryopreservation. Cell Tissue Bank 9, 267–277 (2008). https://doi.org/10.1007/s10561-008-9075-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10561-008-9075-2

Keywords

Navigation