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ORIGINAL RESEARCH article
Front. Endocrinol.
Sec. Reproduction
Volume 15 - 2024 | doi: 10.3389/fendo.2024.1463614
This article is part of the Research TopicFertility Preservation and Restoration in Oncologic and Non-Oncologic PatientsView all 5 articles
Assessment of molecular and morphological dynamics during long-time in vitro cultivation of cryopreserved human ovarian tissue: risk of genetic alterations
Provisionally accepted
Wanxue Wang1
Plamen Todorov Todorov2Evgenia Isachenko1Gohar Rahimi3Markus Merzenich1
Nina Mallmann-Gottschalk1Yang Zhou1
Jilong Yao4Xuemei Li4
Volodimir Isachenko1*- 1University of Cologne, Cologne, Germany
- 2Institute of Biology and Immunology of Reproduction (BAS), Sofia, Sofia City, Bulgaria
- 3Amedes Group GmbH, Hamburg, Hamburg, Germany
- 4Reproductive Medicine Center, Shenzhen Maternity and Child Healthcare Hospital, Shenzhen, Guangdong Province, China
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Background: Cryopreservation of human ovarian tissue is a technology for patients undergoing aggressive anticancer treatments. This technology includes the following stages: saturation by permeable cryoprotectants, freezing, thawing, removal of cryoprotectants, as well as tissues in vitro or in situ culture. Objective: Evaluation of quality of tissue after cryopreservation and in vitro culture with the aim of detection of genetic and molecular changes in cells. Methods: Ovarian tissue was frozen in 6% ethylene glycol and 6% dimethyl sulfoxide with speed of cooling 0.3°C/min and thawed at 100°C. After removal of cryoprotectants tissue fragments were in vitro cultured with the soluble extract of basement membrane protein (Matrigel) 3-D culture system for 7 days. Morphological and functional assessments were conducted using microscopic observation and RNA-Sequencing. Comparative analysis of tissue morphology before and after culture was performed with bioinformatics for gene expression and variant analysis, including functional annotation and study of protein-protein interaction. Results: DNA and RNA analyses after cultivation indicated a rise in gene fusion and alternative splicing events, potentially affecting gene expression and cellular functions. Conclusion: longtime in vitro culture of human ovarian tissue results in substantial changes in its morphology and genetic alteration.
Keywords: human ovarian tissue cryopreservation, in vitro culture, single nucleotide polymorphism (SNP), Insertions and Deletions (InDel), protein kinase inhibitor gamma (PKIG), SE (skipped exon). 2
Received: 12 Jul 2024; Accepted: 27 Aug 2024.
Copyright: © 2024 Wang, Todorov, Isachenko, Rahimi, Merzenich, Mallmann-Gottschalk, Zhou, Yao, Li and Isachenko. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence: Volodimir Isachenko, University of Cologne, Cologne, Germany
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