Skip to main content
SpringerLink
Log in

Application of intra- and extracellular sugars and dimethylsulfoxide to human oocyte cryopreservation

  • ASSISTED REPRODUCTION
  • Published:
Journal of Assisted Reproduction and Genetics Aims and scope Submit manuscript

Abstract

Purpose

Oocyte cryopreservation may avoid many complications of human embryo freezing and provide future fertility for women undergoing cancer therapy. The objective of this study was to explore the application of intra- and extracellular sugars in combination with small amounts of dimethylsulfoxide (DMSO) to human oocyte cryopreservation as an alternative approach.

Methods

Discarded human oocytes that were obtained from IVF patients under informed consent and IRB approval, were cryopreserved by slow cooling to −196°C after being randomly distributed into three groups: (i) DMSO control without intra- and extracellular sugar; (ii) extracellular sugar (raffinose) + DMSO; and (iii) intra- and extracellular sugar (trehalose and raffinose, respectively) + DMSO. Subsequently, all cryopreserved oocytes were thawed rapidly, and their survival was assessed by morphological criteria after 24 h of culture.

Results

A total of 71 oocytes were evaluated in three groups with survival rates of 88.5% (23/26), 68.2% (15/22), and 52.2% (12/23) for intra- and extracellular sugar+DMSO, extracellular sugar+DMSO, and DMSO control groups, respectively.

Conclusion

These results support the use of intra- and extracellular sugars as an alternative approach for cryopreservation of human oocytes.

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

Access this article

Log in via an institution

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

  1. Chen C. Pregnancy after human oocyte cryopreservation. Lancet. 1986;1:884–6. doi:10.1016/S0140-6736(86) 90989-X.

    Article  PubMed  CAS  Google Scholar 

  2. Porcu E, Fabbri R, Seracchioli R, Ciotti PM, Magrini O, Flamigni C. Birth of a healthy female after intracytoplasmic sperm injection of cryopreserved human oocytes. Fertil Steril. 1997;68:724–6. doi:10.1016/S0015-0282(97) 00268-9.

    Article  PubMed  CAS  Google Scholar 

  3. Tucker MJ, Wright G, Morton PC, Massey JB. Birth after cryopreservation of immature oocytes with subsequent in vitro maturation. Fertil Steril. 1998;70:578–9. doi:10.1016/S0015-0282(98) 00205-2.

    Article  PubMed  CAS  Google Scholar 

  4. Yoon TK, Chung HM, Lim JM, Han SY, Ko JJ, Cha KY. Pregnancy and delivery of healthy infants developed from vitrified oocytes in a stimulated in vitro fertilization-embryo transfer program [letter]. Fertil Steril. 2000;74:180–1. doi:10.1016/S0015-0282(00) 00572-0.

    Article  PubMed  CAS  Google Scholar 

  5. Kuleshova L, Gianaroli L, Magli C, Ferraretti A, Trounson A. Birth following vitrification of a small number of human oocytes: case report. Hum Reprod. 1999;14:3077–9. doi:10.1093/humrep/14.12.3077.

    Article  PubMed  CAS  Google Scholar 

  6. Kuwayama M, Vajta G, Kato O, Leibo SP. Highly efficient vitrification method for cryopreservation of human oocytes. Reprod Biomed Online. 2005;11:300–8.

    PubMed  Google Scholar 

  7. Borini A, Lagalla C, Bonu MA, Bianchi V, Flamigni C, Coticchio G. Cumulative pregnancy rates resulting from the use of fresh and frozen oocytes: 7 years’ experience. Reprod Biomed Online. 2006;12:481–6.

    PubMed  Google Scholar 

  8. Boldt J, Tidswell N, Sayers A, Kilani R, Cline D. Human oocyte cryopreservation: 5-year experience with a sodium-depleted slow freezing method. Reprod Biomed Online. 2006;13:96–100.

    Article  PubMed  CAS  Google Scholar 

  9. Antinori M, Licata E, Dani G, Cerusico F, Versaci C, Antinori S. Cryotop vitrification of human oocytes results in high survival rate and healthy deliveries. Reprod Biomed Online. 2007;14:73–9.

    Google Scholar 

  10. Committee AP. Essential elements of informed consent for elective oocyte cryopreservation: a Practice Committee opinion. Fertil Steril. 2007;88:1495–6. doi:10.1016/j.fertnstert.2007.10.009.

    Article  Google Scholar 

  11. Oktay K, Cil AP, Bang H. Efficiency of oocyte cryopreservation: a meta-analysis. Fertil Steril. 2006;86:70–80. doi:10.1016/j.fertnstert.2006.03.017.

    Article  PubMed  Google Scholar 

  12. Eroglu A, Russo MJ, Bieganski R, Fowler A, Cheley S, Bayley H, et al. Intracellular trehalose improves the survival of cryopreserved mammalian cells. Nat Biotechnol. 2000;18:163–7. doi:10.1038/72608.

    Article  PubMed  CAS  Google Scholar 

  13. Eroglu A, Toner M, Toth TL. Beneficial effect of microinjected trehalose on the cryosurvival of human oocytes. Fertil Steril. 2002;77:152–8. doi:10.1016/S0015-0282(01) 02959-4.

    Article  PubMed  Google Scholar 

  14. Eroglu A, Bailey SE, Toner M, Toth TL. Successful cryopreservation of mouse oocytes by using low concentrations of trehalose and dimethylsulfoxide. Biol Reprod. 2009;80:70–8. doi:10.1095/biolreprod.108.070383.

    Article  PubMed  CAS  Google Scholar 

  15. Eroglu A, Lawitts JA, Toner M, Toth TL. Quantitative microinjection of trehalose into mouse oocytes and zygotes, and its effect on development. Cryobiology. 2003;46:121–34. doi:10.1016/S0011-2240(03) 00018-X.

    Article  PubMed  CAS  Google Scholar 

  16. McWilliams RB, Gibbons WE, Leibo SP. Osmotic and physiological responses of mouse zygotes and human oocytes to mono- and disaccharides. Hum Reprod. 1995;10:1163–71.

    PubMed  CAS  Google Scholar 

  17. Crowe JH, Hoekstra FA, Crowe LM. Anhydrobiosis. Annu Rev Physiol. 1992;54:579–99. doi:10.1146/annurev.ph.54.030192.003051.

    Article  PubMed  CAS  Google Scholar 

  18. Potts M. Desiccation tolerance of prokaryotes. Microbiol Rev. 1994;58:755–805.

    PubMed  CAS  Google Scholar 

  19. Beattie GM, Crowe JH, Lopez AD, Cirulli V, Ricordi C, Hayek A. Trehalose: a cryoprotectant that enhances recovery and preserves function of human pancreatic islets after long-term storage. Diabetes. 1997;46:519–23. doi:10.2337/diabetes.46.3.519.

    Article  PubMed  CAS  Google Scholar 

  20. Crowe JH, Crowe LM, Carpenter JF. Preserving dry biomaterials: the water replacement hypothesis, Part I. BioPharm. 1993;28:31.

    Google Scholar 

  21. Crowe JH, Crowe LM, Carpenter JF. Preserving dry biomaterials: the water replacement hypothesis, Part II. BioPharm. 1993;28:40–4.

    Google Scholar 

  22. Crowe JH, Leslie SB, Crowe LM. Is vitrification sufficient to preserve liposomes during freeze-drying? Cryobiology. 1994;31:355–66. doi:10.1006/cryo.1994.1043.

    Article  PubMed  CAS  Google Scholar 

  23. Somero GN. Protons, osmolytes, and fitness of internal milieu for protein function. Am J Physiol. 1986;251:R197–213.

    PubMed  CAS  Google Scholar 

  24. Singer MA, Lindquist S. Thermotolerance in Saccharomyces cerevisiae: the Yin and Yang of trehalose. Trends Biotechnol. 1998;16:460–8. doi:10.1016/S0167-7799(98) 01251-7.

    Article  PubMed  CAS  Google Scholar 

  25. Sola-Penna M, Ferreira-Pereira A, Lemos AP, Meyer-Fernandes JR. Carbohydrate protection of enzyme structure and function against guanidinium chloride treatment depends on the nature of carbohydrate and enzyme. Eur J Biochem. 1997;248:24–9. doi:10.1111/j.1432-1033.1997.00024.x.

    Article  PubMed  CAS  Google Scholar 

  26. Chen Q, Haddad GG. Role of trehalose phosphate synthase and trehalose during hypoxia: from flies to mammals. J Exp Biol. 2004;207:3125–9. doi:10.1242/jeb.01133.

    Article  PubMed  CAS  Google Scholar 

  27. Johnson MH, Pickering SJ. The effect of dimethylsulphoxide on the microtubular system of the mouse oocyte. Development. 1987;100:313–24.

    PubMed  CAS  Google Scholar 

  28. Vanderelst J, Vandenabbeel E, Nerinckx S, Vansteirteghem A. Parthenogenetic Activation Pattern and Microtubular Organization of the Mouse Oocyte after Exposure to 1, 2-Propanediol. Cryobiology. 1992;29:549–62. doi:10.1016/0011-2240(92) 90060-F.

    Article  CAS  Google Scholar 

  29. Eroglu A, Elliott G, Wright DL, Toner M, Toth TL. Progressive elimination of microinjected trehalose during mouse embryonic development. Reprod Biomed Online. 2005;10:503–10.

    Article  PubMed  CAS  Google Scholar 

  30. Huang JYJ, Chen HY, Tan SL, Chian RC. Effects of osmotic stress and cryoprotectant toxicity on mouse oocyte fertilization and subsequent embryonic development in vitro. Cell Preserv Technol. 2006;4:149–60. doi:10.1089/cpt.2006.4.149.

    Article  CAS  Google Scholar 

  31. Larman MG, Katz-Jaffe MG, Sheehan CB, Gardner DK. 1, 2-propanediol and the type of cryopreservation procedure adversely affect mouse oocyte physiology. Hum Reprod. 2007;22:250–9. doi:10.1093/humrep/del319.

    Article  PubMed  CAS  Google Scholar 

  32. Sher G, Keskintepe L, Mukaida T, Keskintepe M, Ginsburg M, Agca Y, et al. Selective vitrification of euploid oocytes markedly improves survival, fertilization and pregnancy-generating potential. Reprod Biomed Online. 2008;17:524–9.

    Article  PubMed  CAS  Google Scholar 

  33. Keskintepe L, Agca Y, Sher G, Keskintepe M, Maassarani G. High survival rate of metaphase II human oocytes after first polar body biopsy and vitrification: determining the effect of previtrification conditions. Fertil Steril. 2008.

  34. Cobo A, Kuwayama M, Perez S, Ruiz A, Pellicer A, Remohi J. Comparison of concomitant outcome achieved with fresh and cryopreserved donor oocytes vitrified by the Cryotop method. Fertil Steril. 2008;89:1657–64. doi:10.1016/j.fertnstert.2007.05.050.

    Article  PubMed  Google Scholar 

Download references

Acknowledgments

The authors wish to thank IVF nurse/coordinator Cynthia Clower and Deanna Nelsen for clinical research assistance. This study was partially supported by grant R01HD049537 from the National Institute of Child Health and Human Development to A.E.

Author information

Authors and Affiliations

  1. Central Georgia Fertility Institute, Department of Obstetrics and Gynecology, Mercer University School of Medicine, 4075 Elnora Dr., Macon, GA, 31210, USA

    Abdelmoneim Younis, David Carnovale & William Butler

  2. Institute of Molecular Medicine and Genetics, Department of Medicine and Department of Obstetrics and Gynecology, Medical College of Georgia, 1120 15th Street, CB-2803, Augusta, GA, 30912, USA

    Ali Eroglu

Authors
  1. Abdelmoneim Younis
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. David Carnovale
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. William Butler
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ali Eroglu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Abdelmoneim Younis or Ali Eroglu.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Younis, A., Carnovale, D., Butler, W. et al. Application of intra- and extracellular sugars and dimethylsulfoxide to human oocyte cryopreservation. J Assist Reprod Genet 26, 341–345 (2009). https://doi.org/10.1007/s10815-009-9316-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10815-009-9316-8

Keywords

Search

Navigation