Skip to main content
SpringerLink
Log in

Cryopreservation of female germ cells and ovarian tissues for fertility preservation

  • Review Article
  • Published:
Reproductive Medicine and Biology

Abstract

To preserve the fertility of patients who undergo chemotherapy and/or radiotherapy, procedures for cryopreservation of female germ cells have been investigated. Cyropreservation methods differ according to follicle stage because the mammalian ovary contains a large number of oocytes at different growth stages. Follicles at very early stages, for example the primordial and primary stages, are usually cryopreserved within ovarian cortical tissue because they need surrounding somatic cells for subsequent development. In contrast, fully-grown oocytes in Graafian follicles are cryopreserved without any other cells at the metaphase II stage. Recently, ultra-rapid cooling was incorporated into cryopreservation procedures for human ovaries. In this review, we describe oocyte freezing, the development of ultra-rapid cooling systems for ovarian tissues, freezing of human ovaries, and ovarian transplantation.

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. Jemal A, Siegel R, Ward E, Hao Y, Xu J, Murray T, Thun MJ. Cancer statistics. CA Cancer J Clin. 2008;58:71–96.

    Article  PubMed  Google Scholar 

  2. Meadows AT. Pediatric cancer survivors: past history and future challenges. Curr Prob Cancer. 2003;27:112–26.

    Article  Google Scholar 

  3. Gadducci A, Cosio S, Genazzani AR. Ovarian function and childbearing issues in breast cancer survivors. Gynecol Endocrinol. 2007;23:625–31.

    Article  PubMed  Google Scholar 

  4. Blumenfeld Z. Preservation of fertility and ovarian function and minimalization of chemotherapy associated gonadotoxicity and premature ovarian failure: the role of inhibin-A and -B as markers. Mol Cell Endocrinol. 2002;187:93–105.

    Article  PubMed  CAS  Google Scholar 

  5. Behringer K, Breuer K, Reineke T, May M, Nogova L, Klimm B, Schmitz T, Wildt L, Diehl V, Engert A. Secondary amenorrhea after Hodgkin’s lymphoma is influenced by age at treatment, stage of disease, chemotherapy regimen and the use of oral contraceptives during therapy: a report from the German Hodgkin’s Lymphoma Study Group. J Clin Oncol. 2005;23:7555–64.

    Article  PubMed  Google Scholar 

  6. Morice P, Juncker L, Rey A, El-Hassan J, Haie-Meder C, Castaigne D. Ovarian transposition for patients with cervical carcinoma treated by radiosurgical combination. Fertil Steril. 2000;74:743–8.

    Article  PubMed  CAS  Google Scholar 

  7. Jeruss JS, Woodruff TK. Preservation of fertility in patients with cancer. N Engl J Med. 2009;26:902–11.

    Article  Google Scholar 

  8. Donnez J, Dolmans MM, Demylle D, Jadoul P, Pirard C, Squifflet J, Martinez-Madrid B, Van Langendonckt A. Livebirth after orthotopic transplantation of cryopreserved ovarian tissue. Lancet. 2004;364:1405–10.

    Article  PubMed  CAS  Google Scholar 

  9. Donnez J. Restoration of ovarian function in orthotopically transplanted cryopreserved ovarian tissue: a pilot experience. Reprod Biomed Online. 2008;16:694–704.

    Article  PubMed  CAS  Google Scholar 

  10. Donnez J, Martinez-Madrid B, Jadoul P, Van Langendonckt A, Demylle D, Dolmans MM. Ovarian tissue cryopreservation and transplantation: a review. Hum Reprod Update. 2006;12:519–35.

    Article  PubMed  Google Scholar 

  11. Meirow D, Levron J, Eldar-Geva T. Pregnancy after transplantation of cryopreserved ovarian tissue in a patient with ovarian failure after chemotherapy. N Engl J Med. 2005;353:318–21.

    Article  PubMed  CAS  Google Scholar 

  12. Demeestrere I, Simon P, Buxant F, Robin V, Fernandez SA, Centner J, Delbaere A, Englert Y. Ovarian function and spontaneous pregnancy after combined heterotopic and orthotopic cryopreserved ovarian tissue transplantation in a patient previously treated with bone marrow transplantation: case report. Hum Reprod. 2006;21:2010–4.

    Article  Google Scholar 

  13. Donnez J, Kim SS, Albertini DF. Proceedings of the first world congress on fertility preservation: executive summary. J Assist Reprod Genet. 2010;27:191–5.

    Article  PubMed  Google Scholar 

  14. Whittingham DG. Survival of mouse embryos after freezing and thawing. Nature. 1971;233:125–6.

    Article  PubMed  CAS  Google Scholar 

  15. Rall W, Fahy G. Ice-free cryopreservation of mouse embryos at −196°C by vitrification. Nature. 1985;313:573–5.

    Article  PubMed  CAS  Google Scholar 

  16. Mazur P. Kinetics of water loss from cells at subzero temperatures and the likelihood of intracellular freezing. J Gen Physiol. 1963;47:347–69.

    Article  PubMed  CAS  Google Scholar 

  17. van Uem JF, Siebzehnrubl ER, Schuh B, Koch R, Trotnow S, Lang N. Birth after cryopreservation of unfertilized oocytes. Lancet. 1987;1:752–3.

    PubMed  Google Scholar 

  18. Gook DA, Osborn SM, Bourne H, Johnston WI. Fertilization of human oocytes following cryopreservation; normal karyotypes and absence of stray chromosomes. Hum Reprod. 1994;9:684–91.

    PubMed  CAS  Google Scholar 

  19. Kazem R, Thompson LA, Srikantharajah A, Laing MA, Hamilton MP, Templeton A. Cryopreservation of human oocytes and fertilization by two techniques: in-vitro fertilization and intracytoplasmic sperm injection. Hum Reprod. 1995;10:2650–4.

    PubMed  CAS  Google Scholar 

  20. 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.

    Article  PubMed  CAS  Google Scholar 

  21. Nawroth F, Kissing K. Pregnancy after intracytoplasmic sperm injection (ICSI) of cryopreserved human oocytes. Acta Obstet Gynecol Scand. 1998;77:462–3.

    Article  PubMed  CAS  Google Scholar 

  22. Young E, Kenny A, Puigdomenech E, Van Thillo G, Tiveron M, Piazza A. Triplet pregnancy after intracytoplasmic sperm injection of cryopreserved oocytes: case report. Fertil Steril. 1998;70:360–1.

    Article  PubMed  CAS  Google Scholar 

  23. Allan J. Re: case report: pregnancy from intracytoplasmic injection of a frozen-thawed oocyte. Aust N Z J Obstet Gynaecol. 2004;44:588.

    Article  PubMed  Google Scholar 

  24. Notrica J, Divita A, Neuspiller F, Arenas G, de Fried EP. Healthy girl born after cryopreservation of gametes and ICSI in a patient with seminoma. Reprod Biomed Online. 2004;9:620–2.

    Article  PubMed  Google Scholar 

  25. Borini A, Sciajno R, Bianchi V, Sereni E, Flamigni C, Coticchio G. Clinical outcome of oocyte cryopreservation after slow cooling with a protocol utilizing a high sucrose concentration. Hum Reprod. 2006;21:512–7.

    Article  PubMed  CAS  Google Scholar 

  26. Levi Setti PE, Albani E, Novara PV, Cesana A, Morreale G. Cryopreservation of supernumerary oocytes in IVF/ICSI cycles. Hum Reprod. 2006;21:370–5.

    Article  PubMed  CAS  Google Scholar 

  27. de Santis L, Cino I, Rabellotti E, Papaleo E, Calzi F, Fusi FM, Brigante C, Ferrari A. Oocyte cryopreservation: clinical outcome of slow-cooling protocols differing in sucrose concentration. Reprod BioMed Online. 2007;14:57–63.

    Article  PubMed  Google Scholar 

  28. Fabbri R, Porcu E, Marsella T, Rocchetta G, Venturoli S, Flamigni C. Human oocyte cryopreservation: new perspectives regarding oocyte survival. Hum Reprod. 2001;16:411–6.

    Article  PubMed  CAS  Google Scholar 

  29. Borini A, Bonu MA, Coticchio G, Bianchi V, Cattoli M, Flamigni C. Pregnancies and births after oocyte cryopreservation. Fertil Steril. 2004;82:601–5.

    Article  PubMed  Google Scholar 

  30. Stachecki JJ, Cohen J, Willadsen SM. Cryopreservation of unfertilized mouse oocytes: the effect of replacing sodium with choline in the freezing medium. Cryobiology. 1998;37:346–54.

    Article  PubMed  CAS  Google Scholar 

  31. Eroglu A, Toner M, Toth TL. Beneficial effect of microinjected trehalose on the cryosurvival of human oocytes. Fertil Steril. 2002;77:152–8.

    Article  PubMed  Google Scholar 

  32. Oktay K, Cil AP, Bang H. Efficiency of oocyte cryopreservation: a meta-analysis. Fertil Steril. 2006;86:70–80.

    Article  PubMed  Google Scholar 

  33. Liebermann J, Tucker MJ. Effect of carrier system on the yield of human oocytes and embryos as assessed by survival and developmental potential after vitrification. Reproduction. 2002;124:483–9.

    Article  PubMed  CAS  Google Scholar 

  34. Vajta G, Nagy ZP. Are programmable freezers still needed in the embryo laboratory? Review on vitrification. Reprod Biomed Online. 2006;12:779–96.

    Article  PubMed  Google Scholar 

  35. Mazur P, Cole KW, Hall WH, Schreuders PD, Mahowald AP. Cryohistological preservation of Drosophila embryos. Science. 1992;258:1932–5.

    Article  PubMed  CAS  Google Scholar 

  36. Martino A, Songsasen N, Leibo SP. Development into blastocysts of bovine oocytes cryopreserved by ultra-rapid cooling. Biol Reprod. 1996;54:1059–69.

    Article  PubMed  CAS  Google Scholar 

  37. Papis K, Shimizu M, Izaike Y. Factors affecting the survivability of bovine oocytes vitrified in droplets. Theriogenology. 2000;54:651–8.

    Article  PubMed  CAS  Google Scholar 

  38. Vajta G, Holm P, Kuwayama M, Booth PJ, Jacobsen H, Greve T, Callesen H. Open pulled straw (OPS) vitrification: a new way to reduce cryoinjuries of bovine ova and embryos. Mol Reprod Dev. 1998;51:53–8.

    Article  PubMed  CAS  Google Scholar 

  39. Hochi S, Hirabayashi M, Hirao M, Kato M, Kobayashi T, Kimura K, Hirasawa K, Leibo SP, Ueda M. Effects of cryopreservation of pronuclear-stage rabbit zygotes on the morphological survival, blastocyst formation, and full-term development after DNA microinjection. Mol Reprod Dev. 2001;60:227–32.

    Article  PubMed  CAS  Google Scholar 

  40. Dinnyés A, Dai Y, Jiang S, Yang X. High developmental rates of vitrified bovine oocytes following parthenogenetic activation, in vitro fertilization, and somatic cell nuclear transfer. Biol Reprod. 2000;63:513–8.

    Article  PubMed  Google Scholar 

  41. Matsumoto H, Jiang JY, Tanaka T, Sasada H, Sato E. Vitrification of large quantities of immature bovine oocytes using nylon mesh. Cryobiology. 2001;42:139–44.

    Article  PubMed  CAS  Google Scholar 

  42. Amo A, Hashimoto S, Nagashima H, Takahashi M, Sasayama N, Morimoto Y. A novel vitrification method using a microfiltration membrane (MFM) enables a simple manipulation of human embryos. Fertil Steril. 2010;94:S116.

    Article  Google Scholar 

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

    Article  PubMed  Google Scholar 

  44. Smith G, Serafini P, Fioravanti J, Yadid I, Coslovsky M, Hassun P, Alegretti JR, Motta EL. Prospective randomized comparison of human oocyte cryopreservation with slow-rate freezing or vitrification. Fertil Steril. 2010;94:2088–95.

    Article  PubMed  Google Scholar 

  45. Cobo A, Kuwayama M, Pérez S, Ruiz A, Pellicer A, Remohí J. Comparison of concomitant outcome achieved with fresh and cryopreserved donor oocytes vitrified by the Cryotop method. Fertil Steril. 2008;89:1657–64.

    Article  PubMed  Google Scholar 

  46. Meryman H. Cryoprotective agents. Cryobiology. 1971;8:173–83.

    Article  PubMed  CAS  Google Scholar 

  47. Mazur P, Leibo SP, Chu EH. A two-factor hypothesis of freezing injury. Evidence from Chinese hamster tissue-culture cells. Exp Cell Res. 1972;71:345–55.

    Article  PubMed  CAS  Google Scholar 

  48. Friedler S, Giudice LC, Lamb EJ. Cryopreservation of embryos and ova. Fertil Steril. 1988;49:743–64.

    PubMed  CAS  Google Scholar 

  49. Larman MG, Sheehan CB, Gardner DK. Calcium-free vitrification reduces cryoprotectant-induced zona pellucida hardening and increases fertilization rates in mouse oocytes. Reproduction. 2006;131:53–61.

    Article  PubMed  CAS  Google Scholar 

  50. Katz-Jaffe MG, Larman MG, Sheehan CB, Gardner D. Exposure of mouse oocytes to 1, 2-propanediol during slow freezing alters the proteome. Fertil Steril. 2008;89:1441–7.

    Article  PubMed  CAS  Google Scholar 

  51. 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.

    Article  PubMed  CAS  Google Scholar 

  52. Fuller B, Paynter S. Fundamentals of cryobiology in reproductive medicine. Reprod Biomed Online. 2004;9:680–91.

    Article  PubMed  Google Scholar 

  53. Hovatta O. Methods for cryopreservation of human ovarian tissue. Reprod Biomed Online. 2005;10:729–34.

    Article  PubMed  Google Scholar 

  54. Deanesly R. Immature rat ovaries grafted after freezing and thawing. J Endocrinol. 1954;11:197–200.

    Article  PubMed  CAS  Google Scholar 

  55. Green SH, Smith AU, Zuckerman S. The number of oocytes in ovarian autografts after freezing and thawing. J Endocrinol. 1956;13:330–4.

    Article  PubMed  CAS  Google Scholar 

  56. Parkes AS. Factors affecting the viability of frozen ovarian tissue. J Endocrinol. 1958;17:337–43.

    Article  PubMed  CAS  Google Scholar 

  57. Parrott DM. The fertility of mice with orthotopic ovarian grafts derived from frozen tissue. J Reprod Fertil. 1960;1:230–41.

    Article  Google Scholar 

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

    PubMed  CAS  Google Scholar 

  59. Hovatta O, Silye R, Krausz T, Abir R, Margara R, Trew G, Lass A, Winston RM. Cryopreservation of human ovarian tissue using dimethylsulphoxide and propanediol–sucrose as cryoprotectants. Hum Reprod. 1996;11:1268–72.

    PubMed  CAS  Google Scholar 

  60. Newton H, Aubard Y, Rutherford A, Sharma V, Gosden R. Low temperature storage and grafting of human ovarian tissue. Hum Reprod. 1996;11:1487–91.

    PubMed  CAS  Google Scholar 

  61. von Wolff M, Donnez J, Hovatta O, Keros V, Maltaris T, Montag M, Salle B, Sonmezer M, Andersen CY. Cryopreservation and autotransplantation of human ovarian tissue prior to cytotoxic therapy—a technique in its infancy but already successful in fertility preservation. Eur J Cancer. 2009;45:1547–53.

    Article  Google Scholar 

  62. Keros V, Xella S, Hultenby K, Pettersson K, Sheikhi M, Volpe A, Hreinsson J, Hovatta O. Vitrification versus controlled-rate freezing in cryopreservation of human ovarian tissue. Hum Reprod. 2009;24:1670–83.

    Article  PubMed  CAS  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  64. Hashimoto S, Suzuki N, Yamanaka M, Hosoi Y, Ishizuka B, Morimoto Y. Effects of vitrification solutions and equilibration times on the morphology of cynomolgus ovarian tissues vitrified ultra-rapidly by direct plunging into liquid nitrogen. Reprod Biomed Online. 2010;21:501–9.

    Article  PubMed  Google Scholar 

  65. Sugimoto M, Maeda S, Manabe N, Miyamoto H. Development of infantile rat ovaries autotransplanted after cryopreservation by vitrification. Theriogenology. 2000;53:1093–103.

    Article  PubMed  CAS  Google Scholar 

  66. Migishima F, Suzuki-Migishima R, Song SY, Kuramochi T, Azuma S, Nishijima M, Yokoyama M. Successful cryopreservation of mouse ovaries by vitrification. Biol Reprod. 2003;68:881–7.

    Article  PubMed  CAS  Google Scholar 

  67. Chen SU, Chien CL, Wu MY, Chen TH, Lai SM, Lin CW, Yang YS. Novel direct cover vitrification for cryopreservation of ovarian tissues increases follicle viability and pregnancy capability in mice. Hum Reprod. 2006;21:2794–800.

    Article  PubMed  Google Scholar 

  68. Wang Y, Xiao Z, Li L, Fan W, Li SW. Novel needle immersed vitrification: a practical and convenient method with potential advantages in mouse and human ovarian tissue cryopreservation. Hum Reprod. 2008;23:2256–65.

    Article  PubMed  CAS  Google Scholar 

  69. Kagawa N, Silber S, Kuwayama M. Successful vitrification of bovine and human ovarian tissue. Reprod BioMed Online. 2009;18:568–77.

    Article  PubMed  Google Scholar 

  70. Isachenko V, Isachenko E, Reinsberg J, Montag M, van der Ven K, Dorn C, Roesing B, van der Ven H. Cryopreservation of human ovarian tissue: comparison of rapid and conventional freezing. Cryobiology. 2007;55:261–8.

    Article  PubMed  CAS  Google Scholar 

  71. Hashimoto S, Suzuki N, Tsuji Y, Yamanaka M, Matsumoto H, Igarashi S, Takae S, Ohta S, Takenoshita M, Yamochi T, Hosoi Y, Ishizuka B, Morimoto Y. Ultra-rapid vitrification supported follicle morphologies of cynomolgus monkeys after freezing compared to conventional vitrification and slow freezing. World Congress on Fertility Preservation; 2009, OC25.

  72. Kristensen SG, Rasmussen A, Byskov AG, Andersen CY. Isolation of pre-antral follicles from human ovarian medulla tissue. Hum Reprod. 2011;26:157–66.

    Article  PubMed  Google Scholar 

  73. Demeestere I, Simon P, Emiliani S, Delbaere A, Englert Y. Fertility preservation: successful transplantation of cryopreserved ovarian tissue in a young patient previously treated for Hodgkin’s disease. Oncologist. 2007;12:1437–42.

    Article  PubMed  Google Scholar 

  74. Andersen CY, Rosendahl M, Byskov AG, Loft A, Ottosen C, Dueholm M, Schmidt K, Andersen AN, Ernst E. Two successful pregnancies following autotransplantation of frozen/thawed ovarian tissue. Hum Reprod. 2008;23:2266–72.

    Article  PubMed  Google Scholar 

  75. Silber SJ, DeRosa M, Pineda J, Lenahan K, Grenia D, Gorman K, Gosden RG. A series of monozygotic twins discordant for ovarian failure: ovary transplantation (cortical versus microvascular) and cryopreservation. Hum Reprod. 2008;23:1531–7.

    Article  PubMed  CAS  Google Scholar 

  76. Oktay K, Buyuk E, Veeck L, Zaninovic N, Xu K, Takeuchi T, Opsahl M, Rosenwaks Z. Embryo development after heterotopic transplantation of cryopreserved ovarian tissue. Lancet. 2004;363:837–40.

    Article  PubMed  Google Scholar 

  77. Oktay K. Spontaneous conceptions and live birth after heterotopic ovarian transplantation: is there a germline stem cell connection? Hum Reprod. 2006;21:1345–8.

    Article  PubMed  Google Scholar 

  78. Rosendahl M, Loft A, Byskov AG, Ziebe S, Schmidt KTL, Andersen AN, Ottosen C, Andersen CY. Biochemical pregnancy after fertilization of an oocyte aspirated from a heterotopic autotransplant of cryopreserved ovarian tissue: case report. Hum Reprod. 2006;21:2006–9.

    Article  PubMed  CAS  Google Scholar 

  79. Lee DM, Yeoman RR, Battaglia DE, Stouffer RL, Zelinski-Wooten MB, Fanton JW, Wolf DP. Live birth after ovarian tissue transplant. Nature. 2004;428:137–8.

    Article  PubMed  CAS  Google Scholar 

  80. Igarashi S, Suzuki N, Hashimoto S, Takae S, Tkenoshita M, Hosoi Y, Morimoto Y, Ishizuka B. Heterotopic autotransplantation of ovarian cortex in cynomolgus monkeys. Hum Cell. 2010;23:26–34.

    PubMed  Google Scholar 

Download references

Acknowledgment

Authors thank Professor Y. Hosoi and Mr T. Yamochi of the Department of Genetic Engineering, Kinki University; Drs S. Igarashi, Y. Okutsu, Y. Sugishita, and M. Ino, Ms M. Inano, M. Hoshina, and S. Tsukamoto of St Marianna University School of Medicine; Dr M. Takenoshita and Mr S. Ohta of Evebioscience; and Ms Y. Tsuji, A. Amo, Mr M. Yamanaka, and H. Matsumoto of IVF Namba Clinic for their kind technical support and helpful comments.

Author information

Authors and Affiliations

  1. IVF Namba Clinic, 1-17-28 Minami-horie, Nishi-ku, Osaka, 5500015, Japan

    Shu Hashimoto & Yoshiharu Morimoto

  2. Department of Obstetrics and Gynecology, St Marianna University School of Medicine, 2-16-1 Sugao, Miyamae-ku, Kawasaki, Kanagawa, 2168511, Japan

    Nao Suzuki & Bunpei Ishizuka

Authors
  1. Shu Hashimoto
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Nao Suzuki
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Bunpei Ishizuka
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Yoshiharu Morimoto
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Shu Hashimoto.

About this article

Cite this article

Hashimoto, S., Suzuki, N., Ishizuka, B. et al. Cryopreservation of female germ cells and ovarian tissues for fertility preservation. Reprod Med Biol 10, 161–169 (2011). https://doi.org/10.1007/s12522-011-0088-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12522-011-0088-3

Keywords

Search

Navigation