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Attempts to improve human ovarian transplantation outcomes of needle-immersed vitrification and slow-freezing by host and graft treatments

  • Ronit AbirEmail author
  • Benjamin Fisch
  • Noa Fisher
  • Nivin Samara
  • Galit Lerer-Serfaty
  • Roei Magen
  • Michal Herman-Edelstein
  • Avi Ben-Haroush
  • Anat Stein
  • Raoul Orvieto
Fertility Preservation

Abstract

Purpose

To investigate if needle-immersed vitrification or slow-freezing yields better implantation results for human ovarian tissue and which method benefits more when combined with the “improvement protocol” of host melatonin treatment and graft incubation with biological glue + vitamin E + vascular endothelial growth factor-A.

Methods

Human ovarian tissue was preserved by needle-immersed vitrification or slow-freezing and transplanted into immunodeficient mice, either untreated (groups A and C, respectively) or treated with the improvement protocol (groups B and D, respectively). Grafted and ungrafted slices were evaluated by follicle counts, apoptosis assay and immunohistochemistry for Ki67 and platelet endothelial cell adhesion molecule (PECAM).

Results

Follicle number in the recovered grafts was limited. The number of atretic follicles was significantly higher after vitrification with/without the improvement protocol and slow-freezing than that after slow-freezing + the improvement protocol. Stroma cell apoptosis was the lowest in the group D. PECAM staining showed a peripheral and diffuse pattern in the group D (mostly normal follicular morphology) and a diffuse pattern in all other groups (few follicles, mostly atretic), with significantly higher diffuse levels in the vitrification groups. Ki67 staining was identified in all normal follicles. Follicles did not survive transplantation in the vitrification groups.

Conclusions

Ovarian sample preparation with slow-freezing + the improvement protocol appears to yield better implantation outcomes than needle-immersed vitrification with/without the improvement protocol. The real quality of frozen tissue can be assessed only after grafting and not after thawing/warming.

Keywords

Human ovarian tissue Needle-immersed vitrification Slow-gradual freezing “Improvement protocol” PECAM Ki67 TUNEL 

Notes

Acknowledgments

The authors are indebted to Ms. G. Ganzach from the Editorial Board of Rabin Medical Center for the English editing. The authors are also grateful to our laboratory technician Carmela Felz for the histological sections.

Compliance with ethical standards

The study was approved by the local institutional ethics committee. Informed consent to donate tissue for the present study was obtained from the patients or the parents of minors.

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Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  • Ronit Abir
    • 1
    • 2
    Email author
  • Benjamin Fisch
    • 1
    • 2
  • Noa Fisher
    • 1
    • 2
  • Nivin Samara
    • 1
    • 2
    • 3
  • Galit Lerer-Serfaty
    • 1
    • 2
  • Roei Magen
    • 1
    • 4
  • Michal Herman-Edelstein
    • 5
  • Avi Ben-Haroush
    • 1
    • 2
  • Anat Stein
    • 1
    • 2
  • Raoul Orvieto
    • 2
    • 6
  1. 1.Infertility and IVF Unit, Beilinson Women HospitalRabin Medical CenterPetach TikvaIsrael
  2. 2.Sackler Faculty of MedicineTel Aviv UniversityTel AvivIsrael
  3. 3.In Vitro Fertilization Unit, Lis Maternity and Women’s HospitalTel Aviv Sourasky Medical CenterTel AvivIsrael
  4. 4.Faculty of Health Sciences, Goldman Medical SchoolBen Gurion University of the NegevBeer ShevaIsrael
  5. 5.Department of Nephrology, Rabin Medical Center, Felsenstein Research Center 49100 and Sackler Faculty of MedicineTel Aviv UniversityTel AvivIsrael
  6. 6.Infertility and IVF Unit, Department of Obstetrics and GynecologyThe Chaim Sheba Medical CenterRamat GanIsrael

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