Skip to main content
SpringerLink
Log in

Efficacy of ovarian tissue cryopreservation in a major European center

  • Fertility Preservation
  • Published:
Journal of Assisted Reproduction and Genetics Aims and scope Submit manuscript

Abstract

Purpose

To evaluate the effect of cryopreservation and thawing of ovarian tissue from oncological patients opting for fertility preservation on ovarian tissue viability.

Methods

In this prospective cohort study, the ovarian tissue viability before and after cryopreservation and thawing was measured for 25 newly diagnosed oncological patients who had their ovarian tissue cryopreserved. Outcome measures were follicle integrity (histology), follicle viability (Calcein viability assay), steroid hormone production (estradiol and progesterone production in vitro) and overall tissue viability (glucose uptake in vitro). This study was conducted at a Cryobank for storage of ovarian tissue in a university hospital.

Results

Cryopreserved/thawed ovarian tissue showed a decreased glucose uptake when compared to tissue that had not been cryopreserved. In addition, a diminished E2 and P4 production was observed after cryopreservation and thawing, despite the fact that numbers of viable follicles as determined by the Calcein viability assay were comparable. Histological examination revealed a higher percentage of degenerated follicles after cryopreservation and thawing.

Conclusions

Ovarian tissue cryopreservation and thawing impairs the viability of ovarian tissue in oncological patients opting for fertility preservation.

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
Fig. 3

Similar content being viewed by others

References

  1. Donnez J, Dolmans MM. Fertility preservation in women. Nat Rev Endocrinol. 2013;9(12):735–49.

    CAS  PubMed  Google Scholar 

  2. Dittrich R, Lotz L, Keck G, Hoffmann I, Mueller A, Beckmann MW, et al. Live birth after ovarian tissue autotransplantation following overnight transportation before cryopreservation. Fertil Steril. 2012;97(2):387–90.

    Article  PubMed  Google Scholar 

  3. Donnez J, Dolmans MM, Demylle D, Jadoul P, Pirard C, Squifflet J, et al. Livebirth after orthotopic transplantation of cryopreserved ovarian tissue. Lancet. 2004;364(9443):1405–10.

    Article  CAS  PubMed  Google Scholar 

  4. Gracia CR, Chang J, Kondapalli L, Prewitt M, Carlson CA, Mattei P, et al. Ovarian tissue cryopreservation for fertility preservation in cancer patients: successful establishment and feasibility of a multidisciplinary collaboration. J Assist Reprod Genet. 2012;29(6):495–502.

    Article  PubMed Central  PubMed  Google Scholar 

  5. Oktay K, Oktem O. Ovarian cryopreservation and transplantation for fertility preservation for medical indications: report of an ongoing experience. Fertil Steril. 2010;93(3):762–8.

    Article  PubMed  Google Scholar 

  6. Poirot C, Abirached F, Prades M, Coussieu C, Bernaudin F, Piver P. Induction of puberty by autograft of cryopreserved ovarian tissue. Lancet. 2012;379(9815):588.

    Article  PubMed  Google Scholar 

  7. Radford JA, Lieberman BA, Brison DR, Smith AR, Critchlow JD, Russell SA, et al. Orthotopic reimplantation of cryopreserved ovarian cortical strips after high-dose chemotherapy for Hodgkin’s lymphoma. Lancet. 2001;357(9263):1172–5.

    Article  CAS  PubMed  Google Scholar 

  8. Sanfilippo S, Canis M, Romero S, Sion B, Dechelotte P, Pouly JL, et al. Quality and functionality of human ovarian tissue after cryopreservation using an original slow freezing procedure. J Assist Reprod Genet. 2013;30(1):25–34.

    Article  PubMed Central  PubMed  Google Scholar 

  9. Schmidt KL, Ernst E, Byskov AG, Nyboe Andersen A, Yding Andersen C. Survival of primordial follicles following prolonged transportation of ovarian tissue prior to cryopreservation. Hum Reprod. 2003;18(12):2654–9.

    Article  CAS  PubMed  Google Scholar 

  10. Kolp LA, Hubayter Z. Autotransplantation of cryopreserved ovarian tissue: a procedure with promise, risks, and a need for a registry. Fertil Steril. 2011;95(6):1879–86.

    Article  PubMed  Google Scholar 

  11. Bedaiwy MA, El-Nashar SA, El Saman AM, Evers JL, Sandadi S, Desai N, et al. Reproductive outcome after transplantation of ovarian tissue: a systematic review. Hum Reprod. 2008;23(12):2709–17.

    Article  PubMed  Google Scholar 

  12. Rosendahl M, Schmidt KT, Ernst E, Rasmussen PE, Loft A, Byskov AG, et al. Cryopreservation of ovarian tissue for a decade in Denmark: a view of the technique. Reprod Biomed Online. 2011;22(2):162–71.

    Article  PubMed  Google Scholar 

  13. Sanchez M, Novella-Maestre E, Teruel J, Ortiz E, Pellicer A. The Valencia Programme for Fertility Preservation. Clin Transl Oncol. 2008;10(7):433–8.

    Article  CAS  PubMed  Google Scholar 

  14. Donnez J, Jadoul P, Pirard C, Hutchings G, Demylle D, Squifflet J, et al. Live birth after transplantation of frozen-thawed ovarian tissue after bilateral oophorectomy for benign disease. Fertil Steril. 2012;98(3):720–5.

    Article  PubMed  Google Scholar 

  15. Jadoul P, Dolmans MM, Donnez J. Fertility preservation in girls during childhood: is it feasible, efficient and safe and to whom should it be proposed? Hum Reprod Update. 2010;16(6):617–30.

    Article  PubMed  Google Scholar 

  16. Feigin E, Abir R, Fisch B, Kravarusic D, Steinberg R, Nitke S, et al. Laparoscopic ovarian tissue preservation in young patients at risk for ovarian failure as a result of chemotherapy/irradiation for primary malignancy. J Pediatr Surg. 2007;42(5):862–4.

    Article  PubMed  Google Scholar 

  17. Poirot CJ, Martelli H, Genestie C, Golmard JL, Valteau-Couanet D, Helardot P, et al. Feasibility of ovarian tissue cryopreservation for prepubertal females with cancer. Pediatr Blood Cancer. 2007;49(1):74–8.

    Article  PubMed  Google Scholar 

  18. Anderson RA, Wallace WH, Baird DT. Ovarian cryopreservation for fertility preservation: indications and outcomes. Reprod (Cambridge, England). 2008;136(6):681–9.

    Article  CAS  Google Scholar 

  19. Revel A, Revel-Vilk S, Aizenman E, Porat-Katz A, Safran A, Ben-Meir A, et al. At what age can human oocytes be obtained? Fertil Steril. 2009;92(2):458–63.

    Article  PubMed  Google Scholar 

  20. Borgstrom B, Hreinsson J, Rasmussen C, Sheikhi M, Fried G, Keros V, et al. Fertility preservation in girls with turner syndrome: prognostic signs of the presence of ovarian follicles. J Clin Endocrinol Metab. 2009;94(1):74–80.

    Article  PubMed  Google Scholar 

  21. Meirow D, Baum M, Yaron R, Levron J, Hardan I, Schiff E, et al. Ovarian tissue cryopreservation in hematologic malignancy: ten years’ experience. Leuk Lymphoma. 2007;48(8):1569–76.

    Article  PubMed  Google Scholar 

  22. Lawrenz B, Jauckus J, Kupka MS, Strowitzki T, von Wolff M. Fertility preservation in >1,000 patients: patient’s characteristics, spectrum, efficacy and risks of applied preservation techniques. Arch Gynecol Obstet. 2011;283(3):651–6.

    Article  PubMed  Google Scholar 

  23. Bastings L, Beerendonk CC, Westphal JR, Massuger LF, Kaal SE, van Leeuwen FE, et al. Autotransplantation of cryopreserved ovarian tissue in cancer survivors and the risk of reintroducing malignancy: a systematic review. Hum Reprod Update. 2013;19(5):483–506.

    Article  CAS  PubMed  Google Scholar 

  24. Donnez J, Dolmans MM, Pellicer A, Diaz-Garcia C, Sanchez Serrano M, Schmidt KT, et al. Restoration of ovarian activity and pregnancy after transplantation of cryopreserved ovarian tissue: a review of 60 cases of reimplantation. Fertil Steril. 2013;99(6):1503–13.

    Article  PubMed  Google Scholar 

  25. Callejo J, Salvador C, Gonzalez-Nunez S, Almeida L, Rodriguez L, Marques L, et al. Live birth in a woman without ovaries after autograft of frozen-thawed ovarian tissue combined with growth factors. J Ovarian Res. 2013;6(1):33.

    Article  PubMed Central  PubMed  Google Scholar 

  26. Schmidt KL, Yding Andersen C, Starup J, Loft A, Byskov AG, Nyboe AA. Orthotopic autotransplantation of cryopreserved ovarian tissue to a woman cured of cancer - follicular growth, steroid production and oocyte retrieval. Reprod Biomed Online. 2004;8(4):448–53.

    Article  Google Scholar 

  27. Campos JR, Rosa-e-Silva JC, Carvalho BR, Vireque AA, Silva-de-Sa MF, Rosa-e-Silva AC. Cryopreservation time does not decrease follicular viability in ovarian tissue frozen for fertility preservation. Clinics (Sao Paulo). 2011;66(12):2093–7.

    Article  Google Scholar 

  28. Chambers EL, Gosden RG, Yap C, Picton HM. In situ identification of follicles in ovarian cortex as a tool for quantifying follicle density, viability and developmental potential in strategies to preserve female fertility. Hum Reprod. 2010;25(10):2559–68.

    Article  CAS  PubMed  Google Scholar 

  29. Hreinsson J, Zhang P, Swahn ML, Hultenby K, Hovatta O. Cryopreservation of follicles in human ovarian cortical tissue. Comparison of serum and human serum albumin in the cryoprotectant solutions. Hum Reprod. 2003;18(11):2420–8.

    Article  CAS  PubMed  Google Scholar 

  30. Isachenko V, Lapidus I, Isachenko E, Krivokharchenko A, Kreienberg R, Woriedh M, et al. Human ovarian tissue vitrification versus conventional freezing: morphological, endocrinological, and molecular biological evaluation. Reprod (Cambridge, England). 2009;138(2):319–27.

    Article  CAS  Google Scholar 

  31. Maltaris T, Dragonas C, Hoffmann I, Mueller A, Beckmann MW, Dittrich R. Simple prediction of the survival of follicles in cryopreserved human ovarian tissue. J Reprod Dev. 2006;52(4):577–82.

    Article  PubMed  Google Scholar 

  32. Oktay K, Nugent D, Newton H, Salha O, Chatterjee P, Gosden RG. Isolation and characterization of primordial follicles from fresh and cryopreserved human ovarian tissue. Fertil Steril. 1997;67(3):481–6.

    Article  CAS  PubMed  Google Scholar 

  33. Rimon E, Cohen T, Dantes A, Hirsh L, Amit A, Lessing JB, et al. Apoptosis in cryopreserved human ovarian tissue obtained from cancer patients: a tool for evaluating cryopreservation utility. Int J Oncol. 2005;27(2):345–53.

    CAS  PubMed  Google Scholar 

  34. Sanfilippo S, Canis M, Ouchchane L, Botchorishvili R, Artonne C, Janny L, et al. Viability assessment of fresh and frozen/thawed isolated human follicles: reliability of two methods (Trypan blue and Calcein AM/ethidium homodimer-1). J Assist Reprod Genet. 2011;28(12):1151–6.

    Article  PubMed Central  PubMed  Google Scholar 

  35. Schubert B, Canis M, Darcha C, Artonne C, Pouly JL, Dechelotte P, et al. Human ovarian tissue from cortex surrounding benign cysts: a model to study ovarian tissue cryopreservation. Hum Reprod. 2005;20(7):1786–92.

    Article  PubMed  Google Scholar 

  36. Fabbri R, Pasquinelli G, Keane D, Magnani V, Paradisi R, Venturoli S. Optimization of protocols for human ovarian tissue cryopreservation with sucrose, 1,2-propanediol and human serum. Reprod Biomed Online. 2010;21(6):819–28.

    Article  CAS  PubMed  Google Scholar 

  37. Isachenko V, Isachenko E, Reinsberg J, Montag M, Braun F, van der Ven H. Cryopreservation of human ovarian tissue: effect of spontaneous and initiated ice formation. Reprod Biomed Online. 2008;16(3):336–45.

    Article  PubMed  Google Scholar 

  38. Marsella T, Sena P, Xella S, La Marca A, Giulini S, De Pol A, et al. Human ovarian tissue cryopreservation: effect of sucrose concentration on morphological features after thawing. Reprod Biomed Online. 2008;16(2):257–67.

    Article  PubMed  Google Scholar 

  39. Demeestere I, Simon P, Emiliani S, Delbaere A, Englert Y. Orthotopic and heterotopic ovarian tissue transplantation. Hum Reprod Update. 2009;15(6):649–65.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  40. Kim SS, Yang HW, Kang HG, Lee HH, Lee HC, Ko DS, et al. Quantitative assessment of ischemic tissue damage in ovarian cortical tissue with or without antioxidant (ascorbic acid) treatment. Fertil Steril. 2004;82(3):679–85.

    Article  CAS  PubMed  Google Scholar 

  41. Gosden RG, Baird DT, Wade JC, Webb R. Restoration of fertility to oophorectomized sheep by ovarian autografts stored at −196° C. Hum Reprod. 1994;9(4):597–603.

    CAS  PubMed  Google Scholar 

  42. Isachenko V, Isachenko E, Reinsberg J, Montag M, van der Ven K, Dorn C, et al. Cryopreservation of human ovarian tissue: comparison of rapid and conventional freezing. Cryobiology. 2007;55(3):261–8.

    Article  CAS  PubMed  Google Scholar 

  43. Isachenko V, Montag M, Isachenko E, van der Ven K, Dorn C, Roesing B, et al. Effective method for in-vitro culture of cryopreserved human ovarian tissue. Reprod Biomed Online. 2006;13(2):228–34.

    Article  PubMed  Google Scholar 

  44. Gougeon A. Dynamics of follicular growth in the human: a model from preliminary results. Hum Reprod. 1986;1(2):81–7.

    CAS  PubMed  Google Scholar 

  45. Liebenthron J, Koster M, Drengner C, Reinsberg J, van der Ven H, Montag M. The impact of culture conditions on early follicle recruitment and growth from human ovarian cortex biopsies in vitro. Fertil Steril. 2013;100(2):483–91.

    Article  CAS  PubMed  Google Scholar 

  46. Gerritse R, Beerendonk CC, Westphal JR, Bastings L, Braat DD, Peek R. Glucose/lactate metabolism of cryopreserved intact bovine ovaries as a novel quantitative marker to assess tissue cryodamage. Reprod Biomed Online. 2011;23(6):755–64.

    Article  CAS  PubMed  Google Scholar 

  47. Ioannou A, Dalle Lucca J, Tsokos GC. Immunopathogenesis of ischemia/reperfusion-associated tissue damage. Clin Immunol. 2011;141(1):3–14.

    Article  CAS  PubMed  Google Scholar 

  48. Jang HR, Rabb H. The innate immune response in ischemic acute kidney injury. Clin Immunol. 2009;130(1):41–50.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  49. van Golen RF, van Gulik TM, Heger M. The sterile immune response during hepatic ischemia/reperfusion. Cytokine Growth Factor Rev. 2012;23(3):69–84.

    Article  PubMed  Google Scholar 

  50. Amorim CA, Dolmans MM, David A, Jaeger J, Vanacker J, Camboni A, et al. Vitrification and xenografting of human ovarian tissue. Fertil Steril. 2012;98(5):1291–8.

    Article  PubMed  Google Scholar 

  51. Fabbri R, Venturoli S, D’Errico A, Iannascoli C, Gabusi E, Valeri B, et al. Ovarian tissue banking and fertility preservation in cancer patients: histological and immunohistochemical evaluation. Gynecol Oncol. 2003;89(2):259–66.

    Article  CAS  PubMed  Google Scholar 

  52. Fauque P, Ben Amor A, Joanne C, Agnani G, Bresson JL, Roux C. Use of trypan blue staining to assess the quality of ovarian cryopreservation. Fertil Steril. 2007;87(5):1200–7.

    Article  PubMed  Google Scholar 

  53. Milenkovic M, Diaz-Garcia C, Wallin A, Brannstrom M. Viability and function of the cryopreserved whole rat ovary: comparison between slow-freezing and vitrification. Fertil Steril. 2012;97(5):1176–82.

    Article  PubMed  Google Scholar 

  54. Isachenko E, Isachenko V, Nawroth F, Rahimi G, Weiss JM. Effect of long-term exposure at suprazero temperatures on activity and viability of human ovarian cortex. Fertil Steril. 2009;91(4 Suppl):1556–9.

    Article  PubMed  Google Scholar 

  55. Bridges PJ, Brusie MA, Fortune JE. Elevated temperature (heat stress) in vitro reduces androstenedione and estradiol and increases progesterone secretion by follicular cells from bovine dominant follicles. Domest Anim Endocrinol. 2005;29(3):508–22.

    Article  CAS  PubMed  Google Scholar 

  56. Gerritse R, Peek R, Sweep F, Thomas C, Braat D, Kremer J, et al. In vitro 17β-oestradiol release as a marker for follicular survival in cryopreserved intact bovine ovaries. Cryo Letters. 2010;31(4):318–28.

    CAS  PubMed  Google Scholar 

  57. Fritz MA, Speroff L. Current concepts of the endocrine characteristics of normal menstrual function: the key to diagnosis and management of menstrual disorders. Clin Obstet Gynecol. 1983;26(3):647–89.

    Article  CAS  PubMed  Google Scholar 

  58. Lund SA, Murdoch J, Van Kirk EA, Murdoch WJ. Mitogenic and antioxidant mechanisms of estradiol action in preovulatory ovine follicles: relevance to luteal function. Biol Reprod. 1999;61(2):388–92.

    Article  CAS  PubMed  Google Scholar 

  59. Ting AY, Yeoman RR, Campos JR, Lawson MS, Mullen SF, Fahy GM, et al. Morphological and functional preservation of pre-antral follicles after vitrification of macaque ovarian tissue in a closed system. Hum Reprod. 2013;28(5):1267–79.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  60. Meirow D, Levron J, Eldar-Geva T, Hardan I, Fridman E, Yemini Z, et al. Monitoring the ovaries after autotransplantation of cryopreserved ovarian tissue: endocrine studies, in vitro fertilization cycles, and live birth. Fertil Steril. 2007;87(2):418 e7- e15.

    Article  Google Scholar 

  61. Luyckx V, Scalercio S, Jadoul P, Amorim CA, Soares M, Donnez J, et al. Evaluation of cryopreserved ovarian tissue from prepubertal patients after long-term xenografting and exogenous stimulation. Fertil Steril. 2013;100(5):1350–7.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

Unconditional funding was received from the Nijmegen Centre of Evidence Based Practice (NCEBP), Oranjestipendium foundation, and Merck Serono.

Conflict of interest

The authors declare that they have no conflict of interest.

Author information

Authors and Affiliations

  1. Department of Obstetrics and Gynecology (791), Radboud university medical center, PO Box 9101, 6500 HB, Nijmegen, The Netherlands

    L. Bastings, J. R. Westphal, C. C. M. Beerendonk, D. D. M. Braat & R. Peek

  2. Department of Gynecological Endocrinology and Reproductive Medicine, University of Bonn, Sigmund-Freud-Strasse 25, 53127, Bonn, Germany

    J. Liebenthron & H. van der Ven

  3. Department of Reproductive Biology, University of Veterinary Medicine Hannover, Foundation, Buenteweg 2, 30559, Hannover, Germany

    B. Meinecke

  4. Department of Gynecological Endocrinology and Fertility disorders, University of Heidelberg, INF 440, 69120, Heidelberg, Germany

    M. Montag

Authors
  1. L. Bastings
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. J. Liebenthron
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. J. R. Westphal
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. C. C. M. Beerendonk
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. H. van der Ven
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. B. Meinecke
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. M. Montag
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. D. D. M. Braat
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. R. Peek
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to L. Bastings.

Additional information

Capsule Efficacy of ovarian tissue cryopreservation.

The first two authors should be regarded as joint first authors.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Bastings, L., Liebenthron, J., Westphal, J.R. et al. Efficacy of ovarian tissue cryopreservation in a major European center. J Assist Reprod Genet 31, 1003–1012 (2014). https://doi.org/10.1007/s10815-014-0239-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10815-014-0239-7

Keywords

Search

Navigation