| Autor: |
Lee CY; Genetic Diagnosis Laboratory, Lee Women's Hospital, Taichung 40652, Taiwan.; Institute of Bioinformatics and Structural Biology, National Tsing Hua University, Hsinchu 30013, Taiwan., Tsai HN; Genetic Diagnosis Laboratory, Lee Women's Hospital, Taichung 40652, Taiwan., Cheng EH; Genetic Diagnosis Laboratory, Lee Women's Hospital, Taichung 40652, Taiwan.; Post Baccalaureate Medicine, National Chung Hsing University, Taichung 40227, Taiwan., Lee TH; Division of Infertility, Lee Women's Hospital, Taichung 40402, Taiwan.; Institute of Medicine, Chung Shan Medical University, Taichung 40201, Taiwan.; Department of Obstetrics and Gynecology, Chung Shan Medical University Hospital, Taichung 40201, Taiwan., Lin PY; Post Baccalaureate Medicine, National Chung Hsing University, Taichung 40227, Taiwan.; Division of Infertility, Lee Women's Hospital, Taichung 40402, Taiwan., Lee MS; Division of Infertility, Lee Women's Hospital, Taichung 40402, Taiwan.; Institute of Medicine, Chung Shan Medical University, Taichung 40201, Taiwan.; Department of Obstetrics and Gynecology, Chung Shan Medical University Hospital, Taichung 40201, Taiwan., Lee CI; Division of Infertility, Lee Women's Hospital, Taichung 40402, Taiwan.; Department of Obstetrics and Gynecology, Chung Shan Medical University Hospital, Taichung 40201, Taiwan.; Department of Obstetrics and Gynecology, School of Medicine, Chung Shan Medical University, Taichung 40201, Taiwan. |
| Abstrakt: |
Blastocyst vitrification has significantly improved embryo transfer methods, leading to higher implantation success rates and better pregnancy outcomes in subsequent frozen embryo transfer cycles. This study aimed to simulate the transcriptional changes caused by vitrifying human blastocysts using mouse blastocysts as a model and to further investigate these changes' effects. Utilizing a human vitrification protocol, we implanted both vitrified and fresh embryos into mice. We observed the implantation success rates and performed transcriptomic analysis on the blastocysts. To validate the results from messenger RNA sequencing, we conducted reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR) to measure the expression levels of specific genes. Based on mRNA profiling, we predicted the microRNAs responsible for the regulation and used qPCR basic microRNA assays for validation. Our observations revealed a higher implantation success rate for vitrified embryos than fresh embryos. Transcriptomic analysis showed that vitrified-warmed blastocysts exhibited differentially expressed genes (DEGs) primarily associated with thermogenesis, chemical carcinogenesis-reactive oxygen species, oxidative phosphorylation, immune response, and MAPK-related signaling pathways. RT-qPCR confirmed increased expression of genes such as Cdk6 and Nfat2 , and decreased expression of genes such as Dkk3 and Mapk10 . Additionally, gene-microRNA interaction predictions and microRNA expression analysis identified twelve microRNAs with expression patterns consistent with the predicted results, suggesting potential roles in uterine epithelial cell adhesion, trophectoderm development, invasive capacity, and immune responses. Our findings suggest that vitrification induces transcriptomic changes in mouse blastocysts, and even small changes in gene expression can enhance implantation success. These results highlight the importance of understanding the molecular mechanisms underlying vitrification to optimize embryo transfer techniques and improve pregnancy outcomes. |
