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The following is a summary of “Assessment of molecular and morphological dynamics during long-time in vitro cultivation of cryopreserved human ovarian tissue: risk of genetic alterations,” published in the May 2025 issue of Frontiers in Endocrinology by Wang et al. 

Cryopreservation of human ovarian tissue, involving saturation with cryoprotectants, freezing, thawing, and removal, followed by in vitro or in situ culture, a technology developed for patients undergoing aggressive anticancer treatments.  

Researchers conducted a retrospective study to evaluate tissue quality after cryopreservation and in vitro culture, aiming to detect genetic and molecular changes in cells.  

They froze ovarian tissue in 6% ethylene glycol and 6% dimethyl sulfoxide at a cooling rate of 0.3^° C/min and thawed it at 100^°C. After removing the cryoprotectants, tissue fragments were cultured in a 3-D system with soluble extract of basement membrane protein (Matrigel) for 7 days. Morphological and functional assessments were executed by microscopic observation and RNA sequencing. A comparative analysis of tissue morphology before and after culture was performed using bioinformatics for gene expression, variant analysis, functional annotation, and protein-protein interaction studies.  

The results showed that DNA and RNA analyses thereafter cultivation revealed increased gene fusion and alternative splicing events, and these changes likely impacted the gene expression and cellular functions.  

Investigators concluded that prolonged in vitro culture of human ovarian tissue resulted in significant morphological changes and genetic alterations. 

Source: frontiersin.org/journals/endocrinology/articles/10.3389/fendo.2024.1463614/full