Skip to main content
Download PDF

Principes de préparation et de congélation des spermatozoïdes testiculaires humains

Principles of testicular sperm preparation and cryopreservation

Andrologie volume 13, pages 128–133 (2003)Cite this article

Resume

L’intérêt et la faisabilité de la cryoconservation des spermatozoïdes testiculaires humains sont maintenant bien démontrés. Toutefois, la réalisation de la congélation de ces spermatozoïdes reste empirique, elle est basée essentiellement sur les techniques utilisées pour les spermatozoïdes de l’éjaculât. Les spermatozoïdes testiculaires sont considérés comme des gamètes encore «immatures» et sont toujours retrouvés en quantité limitée. Une meilleure connaissance de leurs caractéristiques cryobiologiques contribuerait à mieux définir le type d’échantillon à congeler (tissu, suspension cellulaire contenant des spermatozoïdes.…), à faire évoluer les processus de congélation/décongélation et la nature des milieux cryoprotecteurs et à adapter les conditions de stockage. Compte tenu des caractéristiques fonctionnelles de ces spermatozoïdes et du fait qu’ils sont utilisés exclusivement en ICSI, de nouveaux tests d’évaluation de la résistance à la congélation basés notamment sur leur qualité nucléaire devraient être développés.

Abstract

The advantages and feasibility of human testicular spermatozoa cryoconservation for intracytoplasmic sperm injection (ICSI) have now been clearly demonstrated. However, the freezing protocol is based on empirical knowledge obtained from freezing of ejaculated spermatozoa. Testicular spermatozoa may not be fully mature gametes and may also be retrieved in only limited quantities. Little research has been conducted to determine whether they have the same cryobiological requirements as ejaculated spermatozoa. A better understanding of their cryobiological features and assessment of possible subcellular changes after thawing would help to optimize testicular preparations for cryopreservation (whole biopsies, seminiferous tubules, shredded suspension, single spermatozoa, etc.), freezing-thawing procedure, freezing media, and storage. Finally, there is a growing need for welldefined criteria (nuclear quality, etc.) to evaluate the tolerance of testicular spermatozoa to freezing-thawing procedure for ICSI

References

  1. AGCA Y., CRITSER J.K.: Cryopreservation of spermatozoa in assisted reproduction. Sem. Reprod. Med., 2002, 20: 15–23.

    Article  Google Scholar 

  2. CRABBE E., VERHEYEN G., TOURNAYE H. et al.: Freezing of testicular tissue as a minced suspension preserves sperm quality better than whole biopsy freezing when glycerol is used as cryoprotectant. Int. J. Androl., 1999, 22: 43–48.

    Article  PubMed  CAS  Google Scholar 

  3. CRAFT I., TSIRIGOTIS M: Simplified recovery, preparation and cryopreservation of testicular spermatozoa. Hum. Reprod., 1995, 10: 1623–1627.

    PubMed  CAS  Google Scholar 

  4. DE CROO I., VAN DER ELST J., EVERAERT K. et al.: Fertilization, pregnancy and embryo implantation rates after ICSI with fresh or frozen-thawed testicular spermatozoa. Hum. Reprod., 1998, 13: 1893–1897.

    Article  PubMed  Google Scholar 

  5. DEVIREDDY R.V., SWANLUND D.J., ROBERTS K.P. et al.: The effect of extracellular ice and cryoprotective agents on the water permeability parameters of human sperm plasma membrane during freezing. Hum. Reprod., 2000, 15: 1125–1135.

    Article  PubMed  CAS  Google Scholar 

  6. ESTEVES S.C., SHARMA R.K., THOMAS A.J. et al.: Cryopreservation of human spermatozoa with pentoxifylline improves the post-thaw agonist-induced acrosome reaction rate. Hum. Reprod., 1998, 13: 3384–3389.

    Article  PubMed  CAS  Google Scholar 

  7. FRIEDLER S., RAZIEL A., STRASSBURGER D. et al.: Outcome of ICSI using fresh and cryopreserved-thawed testicular spermatozoa in patients with non-mosaïc Klinefelter’s syndrome. Hum. Reprod., 2001, 16: 2616–2620.

    Article  PubMed  CAS  Google Scholar 

  8. GAO D.Y., LIU C., LIU C. et al.: Prevention of osmotic injury to human spermatozoa during addition and removal of glycerol. Hum. Reprod., 1995, 10: 1109–1122.

    PubMed  CAS  Google Scholar 

  9. GIL-SALOM M., ROMERO J., MINGUEZ Y. et al.: Pregnancies after intracytoplasmic sperm injection with cryopreserved testicular spermatozoa. Hum. Reprod., 1996, 11: 1309–1313.

    PubMed  CAS  Google Scholar 

  10. GIRAUD M.N., GRIZARD G., BOUCHER D.: Membrane fluidity predicts the outcome of cryopreservation of human spermatozoa. Hum. Reprod., 2000, 15: 2160–2164.

    Article  PubMed  CAS  Google Scholar 

  11. GRIZARD G., CHEVALIER V., GRIVEAU J.F. et al.: Influence of seminal plasma on cryopreservation of human spermatozoa in a biological material-free medium: study of normal and lowquality semen. Int. J. Androl., 1999, 22: 190–196.

    Article  PubMed  CAS  Google Scholar 

  12. HAIDL G., OPPER C.: Changes in lipids and membrane anisotropy in human spermatozoa during epididymal maturation. Hum. Reprod., 1997, 12: 2720–2723.

    Article  PubMed  CAS  Google Scholar 

  13. HAMMADEH M.E., DEHN C.H., HIPPACH M. et al.: Comparison between computerized slow-stage and static liquid nitrogen vapour freezing methods with respect to the deleterious effect on chromatin and morphology of spermatozoa from fertile and subfertile men. Int. J. Androl., 2001, 24: 66–72.

    Article  PubMed  CAS  Google Scholar 

  14. HAMMERSTEDT R.H., GRAHAM J.K., NOLAN J.P.: Cryopreservation of mammalian sperm: what we ask them to survive. J. Androl., 1990, 11: 73–88.

    PubMed  CAS  Google Scholar 

  15. HAMMITT D.G., BEDIA. E., ROGERS P.R. et al.: Comparison of motility stimulants for cryopreserved human semen. Fertil. Steril., 1989, 52: 495–502.

    PubMed  CAS  Google Scholar 

  16. LE LANNOU D., GRIVEAU J.F., LAURENT M.C. et al.: Azoospermie et microinjection. Andrologie, 2001, 11: 142–148.

    Google Scholar 

  17. LORNAGE J., GISCARD D’eSTAING S., LEJEUNE H. et al.: Les prélèvements testiculaires dans les azoospermies sécrétoires: le point de vue du biologiste de la reproduction sur la lecture du prélèvement. Andrologie, 2001, 11: 127–132.

    Google Scholar 

  18. MARMAR J.: The emergence of specialized procedures for the acquisition, processing and cryopreservation of epididymal and testicular sperm in connection with intracytoplasmic sperm injection. J. Androl., 1998, 19: 517–526.

    PubMed  CAS  Google Scholar 

  19. MORRIS C.J.: A new development in the cryopreservation of sperm. Hum. Fertil., 2002, 5: 23–29.

    Article  Google Scholar 

  20. NAWROTH F., ISACHENKO V., DESSOLE S. et al.: Vitrification of human spermatozoa without cryoprotectants. Cryo Letters, 2002, 23: 93–102.

    PubMed  CAS  Google Scholar 

  21. NIJS M., VAN DER ELST J.: Biological aspects of testicular sperm extraction. Eur. Obstet. Gynecol. Reprod. Biol., 2000, 92: 1–6.

    Article  CAS  Google Scholar 

  22. NOGUEIRA D., BOURGAIN C., VERHEYEN G. et al.: Light and electron microscopic analysis of human testicular spermatozoa and spermatids from frozen and thawed testicular biopsies. Hum. Reprod., 1999, 14: 2041–2049.

    Article  PubMed  CAS  Google Scholar 

  23. ROYERE D., BARTHELEMY C., HAMAMAH S. et al.: Cryopreservation of spermatozoa: a 1996 review. Hum. Reprod. Update, 1996, 2: 553–559.

    Article  PubMed  CAS  Google Scholar 

  24. SAEZ F., MOTTA C., BOUCHER D. et al.: Antioxydant capacity of prostasomes in human semen. Mol. Hum. Reprod., 1998, 4: 667–672.

    Article  PubMed  CAS  Google Scholar 

  25. SCHLEGEL P.N.: Testicular sperm extraction: micro-dissection improves sperm yield with minimal tissue excision. Hum. Reprod., 1999, 14: 131–135.

    Article  PubMed  CAS  Google Scholar 

  26. SCHLEGEL P.N., PALERMO G.D., GOLDSTEIN M. et al.: Testicular sperm extraction with intracytoplasmic sperm injection for non-obstructive azoospermie. Urology, 1997, 49: 435–440.

    Article  PubMed  CAS  Google Scholar 

  27. SCHUBERT B., GRIZARD G., BOUCHER D.: Utilisation des spermatozoïdes testiculaires en ICSI. Intérêt de la culturein vitro. Andrologie, 2001, 11: 9–20.

    Article  Google Scholar 

  28. STANIC P., SONECKI Z., SUCHANEK E.: Effect of pentoxifylline on motility and membrane integrity of cryopreserved human spermatozoa. Int. J. Androl., 2002, 25: 186–190.

    Article  PubMed  CAS  Google Scholar 

  29. STEELE E.K., Mc CLURE N., LEWIS S.: Comparison of the effects of two methods of cryopreservation on testicular sperm DNA. Fertil. Steril., 2000, 74: 450–453.

    Article  PubMed  CAS  Google Scholar 

  30. VERHEYEN G., PLETINCX I., VAN STEIRTEGHEM A.: Effect of freezing method, thawing temperature and post-thaw dilution/washing on motility (CASA) and morphology characteristics of high quality human sperm. Hum. Reprod., 1993, 8: 1678–1684.

    PubMed  CAS  Google Scholar 

  31. WINDT M.L., COETZEE K., KRUGER T.F. et al.: Intracytoplasmic sperm injection with testicular spermatozoa in men with azoospermia. J. Assist. Reprod. Genet., 2002, 19: 53–59.

    Article  PubMed  CAS  Google Scholar 

  32. WOOD S., THOMAS K., SCHNAUFFER K. et al.: Reproductive potential of fresh and cryopreserved epididymal and testicular spermatozoa in consecutive intracytoplasmic sperm injection cycles in the same patients. Fertil. Steril., 2002, 77: 1162–1166.

    Article  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Laboratoire de Biologie du Développement et de la Reproduction-CECOS, Hôtel-Dieu, Boulevard Léon Malfreyt, 63058, Clermont-Ferrand

    Geneviève Grizard, André Force, Laurent Janny & Daniel Boucher

Authors
  1. Geneviève Grizard
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. André Force
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Laurent Janny
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Daniel Boucher
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Geneviève Grizard.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Grizard, G., Force, A., Janny, L. et al. Principes de préparation et de congélation des spermatozoïdes testiculaires humains. Androl. 13, 128–133 (2003). https://doi.org/10.1007/BF03034426

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF03034426

Mots clés

  • congélation
  • décongélation
  • spermatozoïdes testiculaires
  • homme

Key-Words

  • freezing
  • thawing
  • testicular spermatozoa
  • man

Basic and Clinical Andrology

ISSN: 2051-4190