Improving ovarian tissue cryopreservation for oncologic patients: slow freezing versus vitrification, effect of different procedures and devices

Autores de IIS La Fe

• Sonia Herraiz Raya

  Autor

• Edurne Novella Maestre

  Autor

• César Díaz García

  Autor

• Vicente Mirabet Lis

  Autor

• Antonio Pellicer Martínez

  Autor

Participantes ajenos a IIS La Fe

• Rodríguez B
• Sánchez-Serrano M

Grupos

• Medicina Reproductiva

  

  Investigador Principal

  José Morales Roselló

• Regeneración y Trasplante Cardíaco

  

  Leader

  Pilar Sepúlveda Sanchis

Abstract

Objective: To compare slow freezing (SF) with four vitrification techniques (VT) for cryopreservation of ovarian tissue (OT) and to evaluate the best protocol for human OT in a xenograft model. Design: Experimental study. Setting: University hospital. Patient(s): Patients undergoing fertility preservation. Animal(s): Ovariectomized nude mice. Intervention(s): Cryopreservation of bovine OT after SF and four VTs (VT1, VT2, VT3, and VT4) by combining two cryoprotectant vitrification solutions (VS1 and VS2) and two devices (metallic grid and ethyl vinyl acetate bag), after which the cryopreservation of human OT by SF and VT1 and xenograft into nude mice. Main Outcome Measure(s): Follicular densities, proliferation, vascularization, fibrosis, apoptosis, tissue viability. Result(s): The in vitro study in bovine OT showed a lower percentage of quiescent follicles in the SF group but not in the vitrification groups (VT1-VT4). Apoptosis increased and cell proliferation decreased in all the experimental groups except VT1 (20% ethylene glycol, 20% dimethyl sulfoxide, 0.5 M sucrose, and 20% synthetic serum substitute in HEPES-buffered M199 culture media with Cryotissue metallic grids). Tissue viability was diminished in VT3, and the SF-xenografted human samples showed reduced primordial and secondary densities and unbalanced follicular populations when compared with fresh and VT1 tissue. Conclusion(s): VT1 offers similar conditions to fresh tissue for follicular density, proliferation, viability, and cell death and preserves a larger population of quiescent follicles than SF after transplantation, thus ensuring the maintenance of graft potential fertility. (C) 2014 by American Society for Reproductive Medicine.

Keywords

• Fertility preservation; ovarian cortex; slow freezing; vitrification