AUTHOR=Wang Wanxue , Todorov Plamen , Isachenko Evgenia , Rahimi Gohar , Merzenich Markus , Mallmann-Gottschalk Nina , Zhou Yang , Yao Jilong , Li Xuemei , Isachenko Volodimir 

TITLE=Assessment of molecular and morphological dynamics during long-time in vitro cultivation of cryopreserved human ovarian tissue: risk of genetic alterations

JOURNAL=Frontiers in Endocrinology

VOLUME=Volume 15 - 2024

YEAR=2025

URL=https://www.frontiersin.org/journals/endocrinology/articles/10.3389/fendo.2024.1463614

DOI=10.3389/fendo.2024.1463614

ISSN=1664-2392

ABSTRACT=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.