Successful vitrification and autografting of baboon (Papio anubis) ovarian tissue

Hum Reprod. 2013 Aug;28(8):2146-56. doi: 10.1093/humrep/det103. Epub 2013 Apr 16.

Abstract

Study question: Can a vitrification protocol using an ethylene glycol/dimethyl sulphoxide-based solution and a cryopin successfully cryopreserve baboon ovarian tissue?

Summary answer: Our results show that baboon ovarian tissue can be successfully cryopreserved with our vitrification protocol.

What is known already: Non-human primates have already been used as an animal model to test vitrification protocols for human ovarian tissue cryopreservation.

Study design, size, duration: Ovarian biopsies from five adult baboons were vitrified, warmed and autografted for 5 months.

Participants/materials, setting, methods: After grafting, follicle survival, growth and function and also the quality of stromal tissue were assessed histologically and by immunohistochemistry. The influence of the vitrification procedure on the cooling rate was evaluated by a computer model.

Main results: After vitrification, warming and long-term grafting, follicles were able to grow and maintain their function, as illustrated by Ki67, anti-Müllerian hormone (AMH) and growth differentiation factor-9 (GDF-9) immunostaining. Corpora lutea were also observed, evidencing successful ovulation in all the animals. Stromal tissue quality did not appear to be negatively affected by our cryopreservation procedure, as demonstrated by vascularization and proportions of fibrotic areas, which were similar to those found in fresh ungrafted ovarian tissue.

Limitations, reasons for caution: Despite our promising findings, before applying this technique in a clinical setting, we need to validate it by achieving pregnancies.

Wider implications of the findings: In addition to encouraging results obtained with our vitrification procedure for non-human ovarian tissue, this study also showed, for the first time, expression of AMH and GDF-9 in ovarian follicles.

Study funding/competing interest(s): This study was supported by grants from the Fonds National de la Recherche Scientifique de Belgique (grant Télévie No. 7.4507.10, grant 3.4.590.08 awarded to Marie-Madeleine Dolmans), Fonds Spéciaux de Recherche, Fondation St Luc, Foundation Against Cancer, and Department of Mechanical Engineering at Louisiana State University (support to Ram Devireddy), and donations from Mr Pietro Ferrero, Baron Frère and Viscount Philippe de Spoelberch. None of the authors has any competing interests to declare.

Keywords: autografting; baboon; ovarian tissue; pre-antral follicles; vitrification.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Anti-Mullerian Hormone / metabolism
  • Cell Proliferation
  • Corpus Luteum / physiology
  • Cryopreservation / methods
  • Cryopreservation / veterinary*
  • Female
  • Growth Differentiation Factor 9 / metabolism
  • Immunohistochemistry
  • Ovarian Follicle / metabolism
  • Ovarian Follicle / physiology
  • Ovary / cytology
  • Ovary / transplantation*
  • Papio*
  • Stromal Cells / cytology
  • Transplantation, Autologous / methods
  • Transplantation, Autologous / veterinary

Substances

  • Growth Differentiation Factor 9
  • Anti-Mullerian Hormone