Transcriptomic Analysis of Vitrified-Warmed Vs. Fresh Mouse Blastocysts: Cryo-Induced Physiological Mechanisms and Implantation Impact

Overview

Journal Int J Mol Sci

Publisher MDPI

Specialties Biochemistry
Chemistry
Molecular Biology

Date 2024 Aug 29

PMID 39201343

Authors

Chi-Ying Lee

Han-Ni Tsai

En-Hui Cheng

Tsung-Hsien Lee

Pin-Yao Lin

Maw-Sheng Lee

Chun-I Lee

Affiliations

Soon will be listed here.

Abstract

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 and , and decreased expression of genes such as and . 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.

Citing Articles

Vitrification affects the post-implantation development of mouse embryos by inducing DNA damage and epigenetic modifications.

Chen Y, Zhu H, Guo F, Wang L, Zhang W, Liu R Clin Epigenetics. 2025; 17(1):20.

PMID: 39920865 PMC: 11806629. DOI: 10.1186/s13148-025-01826-y.

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