title: Good preservation of stromal cells and no apoptosis in human ovarian tissue after vitrification.
abstract: The aim of this study was to develop a vitrification procedure for human ovarian tissue cryopreservation in order to better preserve the ovarian tissue. Large size samples of ovarian tissue retrieved from 15 female-to-male transgender subjects (18-38 years) were vitrified using two solutions (containing propylene glycol, ethylene glycol, and sucrose at different concentrations) in an open system. Light microscopy, transmission electron microscopy, and TUNEL assay were applied to evaluate the efficiency of the vitrification protocol. After vitrification/warming, light microscopy showed oocyte nucleus with slightly thickened chromatin and irregular shape, while granulosa and stromal cells appeared well preserved. Transmission electron microscopy showed oocytes with slightly irregular nuclear shape and finely dispersed chromatin. Clear vacuoles and alterations in cellular organelles were seen in the oocyte cytoplasm. Stromal cells had a moderately dispersed chromatin and homogeneous cytoplasm with slight vacuolization. TUNEL assay revealed the lack of apoptosis induction by vitrification in all ovarian cell types. In conclusion after vitrification/warming the stromal compartment maintained morphological and ultrastructural features similar to fresh tissue, while the oocyte cytoplasm was slightly damaged. Although these data are encouraging, further studies are necessary and essential to optimize vitrification procedure.
authors.affiliation: Gynecology and Pathophysiology of Human Reproduction Unit, DIMEC, S.Orsola-Malpighi Hospital, University of Bologna, Via Massarenti 13, 40138 Bologna, Italy.
authors.forename: Raffaella
authors.identifier: 0000-0002-3366-3113
authors.initials: R
authors.lastname: Fabbri
authors.affiliation: Gynecology and Pathophysiology of Human Reproduction Unit, DIMEC, S.Orsola-Malpighi Hospital, University of Bologna, Via Massarenti 13, 40138 Bologna, Italy.
authors.forename: Rossella
authors.initials: R
authors.lastname: Vicenti
authors.affiliation: Gynecology and Pathophysiology of Human Reproduction Unit, DIMEC, S.Orsola-Malpighi Hospital, University of Bologna, Via Massarenti 13, 40138 Bologna, Italy.
authors.forename: Maria
authors.identifier: 0000-0002-5651-1083
authors.initials: M
authors.lastname: Macciocca
authors.affiliation: Clinical Pathology, DIMES, S.Orsola-Malpighi Hospital, University of Bologna, Via Massarenti 9, 40138 Bologna, Italy.
authors.forename: Gianandrea
authors.identifier: 0000-0003-1506-1829
authors.initials: G
authors.lastname: Pasquinelli
authors.affiliation: Gynecology and Pathophysiology of Human Reproduction Unit, DIMEC, S.Orsola-Malpighi Hospital, University of Bologna, Via Massarenti 13, 40138 Bologna, Italy.
authors.forename: Roberto
authors.identifier: 0000-0002-0659-452X
authors.initials: R
authors.lastname: Paradisi
authors.affiliation: Gynecology and Pathophysiology of Human Reproduction Unit, DIMEC, S.Orsola-Malpighi Hospital, University of Bologna, Via Massarenti 13, 40138 Bologna, Italy.
authors.forename: Cesare
authors.initials: C
authors.lastname: Battaglia
authors.affiliation: Veterinary Clinics and Animal Productions Unit, Department of Emergency and Organ Transplantation (DETO), University of Bari Aldo Moro, Valenzano, 70010 Bari, Italy.
authors.forename: Nicola Antonio
authors.identifier: 0000-0001-6853-2757
authors.initials: NA
authors.lastname: Martino
authors.affiliation: Gynecology and Pathophysiology of Human Reproduction Unit, DIMEC, S.Orsola-Malpighi Hospital, University of Bologna, Via Massarenti 13, 40138 Bologna, Italy.
authors.forename: Stefano
authors.initials: S
authors.lastname: Venturoli
mesh_terms: D017209:Apoptosis / Q000235:genetics; D015925:Cryopreservation / Q000379:methods*; D019855:Ethylene Glycol / Q000494:pharmacology; D005260:Female; D006801:Humans; D006080:Ovarian Follicle / Q000166:cytology*; D010053:Ovary / Q000166:cytology*; D017154:Stromal Cells / Q000166:cytology*; D058989:Vitrification
chemical_list: D019855:Ethylene Glycol