GEOapplication/xmlftp://ftp.ncbi.nlm.nih.gov/geo/series/GSE334nnn/GSE334372/primaryOK200GenomicsHomo sapiensNon-coding RNA profiling by high throughput sequencinghttps://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE334372GEOGSEfalseAltered miRNA Expression Profile in Human Sperm and Seminal Plasma Following Cryopreservation-Thawing: A Small RNA Sequencing StudyCryopreservation of human semen is widely used in assisted reproductive technology (ART) and male fertility preservation, but inevitably induces cryodamage to spermatozoa. Small non-coding RNAs, particularly microRNAs (miRNAs), play critical roles in post-transcriptional gene regulation and may reflect cellular responses to cryopreservation stress. Seminal plasma, the natural fluid environment of spermatozoa, contains abundant miRNAs that may also reflect post-thaw biological responses. However, the impact of freeze-thawing on miRNA expression in both sperm and seminal plasma remains poorly understood. In this study, we performed small RNA sequencing on sperm and seminal plasma samples from healthy normozoospermic donors. For sperm, 5 individual donors provided paired fresh and post-thaw samples (A1-A5: post-thaw; B1-B5: fresh). For seminal plasma, 30 donors were randomly divided into 3 groups (n=10 per group), pooled, and split into fresh control (D3, D4, D5) and cryopreservation-thawing (C3, C4, C5) groups. In sperm, cryopreservation-thawing induced [X] differentially expressed miRNAs ([Y] upregulated, [Z] downregulated). In seminal plasma, we identified 15 differentially expressed miRNAs (9 upregulated, 6 downregulated). GO and KEGG analyses revealed enrichment in fatty acid degradation, Toll-like receptor signaling, and oxidative phosphorylation pathways in both sample types. Our findings provide a comprehensive resource for understanding the molecular mechanisms of sperm cryodamage and developing biomarker-based tools for post-thaw semen quality assessment in ART settings.2026/06/30GSE334372GSM9787069GSM9787068GSM9787076GSM9787065GSM9787075GSM9787064GSM9787067GSM9787078GSM9787066GSM9787077GSM9787072GSM9787071GSM9787063GSM9787074GSM9787073GSM978707024676334372Homo sapiens