Positive Effect of Elevated Thawing Rate for Cryopreservation of Human Ovarian Tissue: Transcriptomic Analysis of Fresh and Cryopreserved Cells

Int J Mol Sci. 2024 Dec 23;25(24):13747. doi: 10.3390/ijms252413747.

Abstract

Ovarian tissue cryopreservation has been gradually applied. It is essential to elucidate the differences between cryopreserved and fresh ovarian tissue and to refine cryopreservation protocols for improved outcomes. To explore the transcriptomic differences between fresh ovarian tissue and tissue cryopreserved with an elevated thawing rate. Ovarian tissue samples were collected and cryopreserved (frozen and thawed) following RNA sequencing and histological evaluation. Three groups were formed: fresh tissue (Group 1), frozen tissue after quick thawing at 100 °C (Group 2), and frozen tissue after slow thawing at 37 °C (Group 3). KEGG analysis showed that in comparison with Group 1, DEGs in Group 2 were mainly enriched in the cortisol synthesis and ovarian steroidogenesis pathways, and DEGs in the cells of Group 3 were mainly enriched in the ovarian steroidogenesis pathway. GO analysis showed that compared to cells of Group 2, DEGs in Group 3 were primarily enriched in the SRP-dependent co-translational protein targeting pathway and co-translational protein targeting to the membrane. The results were formulated with a minimal difference in the histological evaluation of cells after quick and slow thawed tissue. Cryopreservation of ovarian tissue by the described method does not decrease follicle production but downregulates the ovarian steroidogenesis pathway, reducing estrogen and progesterone secretion. The quick thawing of ovarian tissue increases the proliferation and apoptosis pathways of cells.

Keywords: Kyoto encyclopedia of genes and genomes (KEGG); RNA sequencing; cryopreservation; differentially expressed genes (DEGs); gene ontology (GO); human ovarian tissue; thawing; transcriptomics.

MeSH terms

  • Adult
  • Cryopreservation* / methods
  • Female
  • Gene Expression Profiling* / methods
  • Humans
  • Ovary* / metabolism
  • Transcriptome*

Grants and funding

This research was funded by the China Scholarship Council for Qingduo Kong (No. 202308230130) and Cheng Pei (No. 202208080057). It was also supported by the Bulgarian National Science Fund (Grant KP-06-H81/9).