000160919 001__ 160919
000160919 005__ 20251017144624.0
000160919 0247_ $$2doi$$a10.1186/s13048-025-01593-7
000160919 0248_ $$2sideral$$a144089
000160919 037__ $$aART-2025-144089
000160919 041__ $$aeng
000160919 100__ $$aMarco, Alicia
000160919 245__ $$aTreatment with trypLE before freezing improves thawing integrity and functionality of sheep ovarian tissue
000160919 260__ $$c2025
000160919 5060_ $$aAccess copy available to the general public$$fUnrestricted
000160919 5203_ $$aObjective
To study innovative approaches to ovarian tissue cryopreservation, a critical issue for fertility preservation in pediatric cancer patients. Despite historical attempts, recent advances in cancer treatment have underscored the urgent need for more effective and reliable ovarian tissue cryopreservation methods. Our research aims to evaluate if decreasing the rigidity of stroma before cryopreservation by investigating pre-treatments with enzymes can enhance the quality of ovarian tissue post-thawing.
Design
Our research evaluated the use of five commonly used enzymes to disaggregate tissue (trypLE, collagenase, dispase, accutase and hyaluronidase) before freezing ovarian tissue to decrease rigidity and facilitate cryopreservation. Sheep ovaries, with high similarity to human ovaries, were used as an animal model. Tissue structure, cell proliferation, apoptosis and viability were assessed before and after thawing.
Results
Our findings showed that enzymatic treatment with trypLE before freezing offered immediate benefits post-thawing with the highest viability values and percentage of intact follicles. A decrease in viability was observed after thawing and culturing the samples. The pretreatment with accutase damaged the tissue severely with also the lowest viability values. Ki67-positive follicles and stromal cells were observed in fresh samples, but only trypLE and hyaluronidase maintained Ki67-positive antral follicles after 2 days culture. Besides, only trypLE maintained all follicles negative to caspase-3 after thawing, and 7 days after culture primordial follicles were apoptotic in all treatments apart from trypLE.
Conclusion
our findings suggest that trypLE pretreatment could provide a beneficial approach for maintaining the functions and viability of cryopreserved ovaries after thawing. Further research is needed to fully understand their impact and optimize cryopreservation protocols in this important clinical context.
000160919 540__ $$9info:eu-repo/semantics/openAccess$$aby-nc-nd$$uhttps://creativecommons.org/licenses/by-nc-nd/4.0/deed.es
000160919 655_4 $$ainfo:eu-repo/semantics/article$$vinfo:eu-repo/semantics/publishedVersion
000160919 700__ $$aGargallo, Marta$$uUniversidad de Zaragoza
000160919 700__ $$0(orcid)0000-0002-8666-622X$$aCiriza, Jesús$$uUniversidad de Zaragoza
000160919 700__ $$aRoyo-Cañas, María
000160919 700__ $$aIbáñez-Deler, Alejandro
000160919 700__ $$0(orcid)0000-0002-1075-8267$$aRemacha, Ana Rosa
000160919 700__ $$aFons-Contreras, María
000160919 700__ $$aMalo, Clara$$uUniversidad de Zaragoza
000160919 7102_ $$11003$$2443$$aUniversidad de Zaragoza$$bDpto. Anatom.Histolog.Humanas$$cArea Histología
000160919 7102_ $$11009$$2617$$aUniversidad de Zaragoza$$bDpto. Patología Animal$$cÁrea Medicina y Cirugía Animal
000160919 773__ $$g18, 94 (2025), [12 pp.]$$tJournal of ovarian research$$x1757-2215
000160919 8564_ $$s2790292$$uhttps://zaguan.unizar.es/record/160919/files/texto_completo.pdf$$yVersión publicada
000160919 8564_ $$s2021316$$uhttps://zaguan.unizar.es/record/160919/files/texto_completo.jpg?subformat=icon$$xicon$$yVersión publicada
000160919 909CO $$ooai:zaguan.unizar.es:160919$$particulos$$pdriver
000160919 951__ $$a2025-10-17-14:23:20
000160919 980__ $$aARTICLE