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. 2022 Mar 11;23(6):3047.
doi: 10.3390/ijms23063047.

Cryoprotectants-Free Vitrification and Conventional Freezing of Human Spermatozoa: A Comparative Transcript Profiling

Mengying Wang  1 , Plamen Todorov  2 , Wanxue Wang  1 , Evgenia Isachenko  1 , Gohar Rahimi  1 , Peter Mallmann  1 , Vladimir Isachenko  1
Affiliations

Cryoprotectants-Free Vitrification and Conventional Freezing of Human Spermatozoa: A Comparative Transcript Profiling

Mengying Wang et al. Int J Mol Sci. .

Abstract

Introduction: Spermatozoa cryopreservation is an important technique to preserve fertility for males. This study aimed at exploring the stability of epigenetics information in human spermatozoa, manipulated by two different technologies, freezing and vitrification.

Methods: Spermatozoa samples were distributed into three groups: 1. Fresh spermatozoa (control group), 2. Frozen spermatozoa, 3. Vitrified spermatozoa. Epigenetic differences of fresh and cryopreserved spermatozoa were evaluated using high-throughput RNA sequencing.

Results: Differentially expressed genes (DEGs) in frozen (1103 genes) and vitrified (333 genes) spermatozoa were evaluated. The bioinformatical analysis identified 8 and 15 significant pathways in groups of frozen and vitrified spermatozoa, respectively. The majority of these pathways are most relevant to immune and infectious diseases. The DEGs of the fertilization process are not detected during vitrification. The freezing process induces more down-regulation of genes and is relevant to apoptosis changes and immune response.

Conclusion: Cryopreservation of human spermatozoa is an epigenetically safe method for male fertility preservation. Cryoprotectant-free vitrification can induce more minor biological changes in human spermatozoa, in comparison with conventional freezing.

Keywords: DEG; cryopreservation; differentially expressed genes; freezing; human spermatozoa; transcriptome stability; vitrification.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
The differential expression genes level in Slow freezing (A) and Vitrification (B) groups.
Figure 2
Figure 2
(A) Significant Gene ontology terms in Slow freezing group (biological process); (B) Significant Gene ontology terms in Vitrification group (biological process); (C) Bubble chart of significant KEGG pathways in Slow freezing group; (D) Bubble chart of significant KEGG pathway in Vitrification group.
Figure 3
Figure 3
(A) Protein–protein interaction in slow freezing group after GSEA enrichment; (B) Protein–protein interaction in vitrification group after GSEA enrichment.
Figure 4
Figure 4
Flow chart of experimental methodology.
See this image and copyright information in PMC

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