Clinical trial NCT06724471: Preservation of Women's Fertility: Evaluation of Innovative Methods for Ovarian Tissue Cryopreservation
Official title: Preservation of Women's Fertility: Evaluation of Innovative Methods for Ovarian Tissue Cryopreservation
Study type: OBSERVATIONAL
Overall status: RECRUITING
Conditions: Infertility, Female
Keywords: Human ovarian tissue
Cryopreservation
Fertility Preservation
Organotypic culture
Female infertility
Lead sponsor: University Hospital, Clermont-Ferrand
Sponsor class: OTHER
Collaborators: name: CryoBioSystem
name: IMoST UMR 1240 Inserm-Université Clermont Auvergne
First posted: 2024-12-09
Enrollment: 30
Sex: FEMALE
Minimum age (years): 18
Maximum age (years): 37
Standard ages: ADULT
Accepts healthy volunteers: no
Observational model: COHORT
Time perspective: PROSPECTIVE
Sampling method: NON_PROBABILITY_SAMPLE
Study population: Adult woman scheduled for benign ovarian cyst resection in the University Hospital Center of Clermont-Ferrand.

Brief summary: The recent innovations in cancer diagnosis and therapy have improved the five-year survival for patients. However, anticancer treatments may impair patient fertility; therefore fertility preservation is recommended before therapy initiation. The sole method for preserving young prepubescent girls' fertility, which can also be used for pubescent women, is ovarian tissue cryopreservation (OTC) with later auto-transplantation. Although 200 births have been reported worldwide after OTC and transplantation, significant improvements are required. Indeed, freezing and thawing protocols vary according to laboratories (media, cryoprotectants, freezing curve, etc...) and selection criteria are not justified. In addition, most of the laboratories use unsafe devices (e.g. screw cap cryovials) for OTC, exposing ovarian tissues to biological hazards during sample storage in nitrogen tanks. To eliminate these risks, novel "high security" devices have been commercialized (welded cryotubes). However, while thermal welding could alter tissue quality, the functionality of the human ovarian tissue frozen with these innovative devices has not yet been evaluated. The objectives of our study are i) to optimize the freezing and the thawing protocols for human OTC according to thermodynamic properties of the freezing medium and the type of device (welded or screwed cryotube) and ii) to determine if the type of cryotube influences the quality of human ovarian tissue.

This project will enable to reach a better understanding of the impact of freezing on ovarian tissue functionality, as well as the implementation of an optimal protocol for OTC within the ART laboratory of Clermont-Ferrand hospital to optimize patient care.

Detailed description: The aim of this project is to determine whether the type of cryogenic tube (with screw caps or thermosoldered) influences the quality and functionality of ovarian tissue after cryopreservation. First, a characterization of the thermodynamic properties of the freezing medium using a differential scanning calorimeter (Diamond DSC, PerkinElmer) will be carried out to optimize freezing and thawing protocols. This analysis will define important parameters such as crystallization temperature (Tc), end-of-melting temperature (Tm), and transition temperatures (Tg'1). Once freezing and thawing protocols are optimized, we will use human ovarian cortex surrounding benign cysts, a model previously validated by our laboratory, to determine whether the type of cryotube influences the quality of human ovarian tissue. These ovarian cortical samples usually destroyed in clinics share similar characteristics with normal ovarian cortex. Ovarian tissue will be cut into 1mm3 fragments and divided into three groups: fresh ovarian cortex (group1, control), ovarian cortex cryopreserved in thermosoldered cryogenic tubes (group 2) or in screw cap (group 3). We will assess ovarian tissue quality immediately after resection for group 1 or after thawing for group 2 and 3 by analyzing follicle density and morphology (HES staining) and proliferation/apoptosis balance (Immunohistochemistry (IHC) for KI67 and cleaved caspase 3). To assess the functionality of the ovarian tissue after thawing, ovarian cortex fragments will be in vitro cultured. We will analyze i) follicle density, type and morphology (HES staining), ii) the proliferation/apoptosis balance (IHC for KI67 and cleaved caspase 3), iii) the expression levels of major folliculogenesis regulators (IHC for GDF-9) and iv) the levels of folliculogenesis-associated hormones (AMH, estrogen) secreted in culture medium.

Eligibility criteria: Inclusion Criteria:

* Adult woman scheduled for benign ovarian cyst resection (dermoid, functional, mucinous or serous cysts)
* Below 37 years old and above 18 years old
* Capable of giving written informed consent to participate in the research study
* Affiliated to social welfare service

Exclusion Criteria:

* Women above 37 years old and below 18 years old
* Polycystic ovary syndrome
* Diminished ovarian reserve
* Severe endometriosis
* Malignant and endometrial cysts

Primary outcomes:
measure: Post-thawing Ovarian Follicle morphology
timeFrame: 24 months
Secondary outcomes:
measure: Post-thawing Ovarian Follicle density
timeFrame: 24 months
measure: Post-thawing Ovarian Follicle proliferation/apoptosis
timeFrame: 24 months
measure: Ovarian Follicle stage after organotypic culture
timeFrame: 36 months
measure: Ovarian Follicle morphology after organotypic culture
timeFrame: 36 months
measure: GDF-9 expression after organotypic culture
timeFrame: 36 months
measure: Proliferation/apoptosis after organotypic culture
timeFrame: 36 months
measure: hormone secretion after organotypic culture
timeFrame: 36 months
measure: Thermodynamic properties of freezing medium
timeFrame: 12 months
Other outcomes:
Locations:
city: Clermont-Ferrand
country: France
facility: University Hospital
Study officials:
affiliation: University Hospital, Clermont-Ferrand
name: Florence Brugnon, MD, PhD
role: PRINCIPAL_INVESTIGATOR
References: