Abstract
Although vitrification has been widely applied in assisted reproductive technology, it is unknown whether storage time has any impact on the mRNA and lncRNA expression profiles in human embryos. Eleven women (aged 23–35 years) who had undergone in vitro fertilization treatment were recruited for this study. The transcriptomes of 3 fresh eight-cell embryos and 8 surviving vitrified-warmed eight-cell embryos (4 embryos were cryostored for 3 years, and the others were cryostored for 8 years) were analyzed through single-cell RNA-Seq. No differentially expressed mRNAs or lncRNAs were identified between the 3-years group and 8-years group. A total of 128 mRNAs and 365 lncRNAs were differentially expressed in the 8 vitrified-warmed embryos compared with the fresh embryos. The vitrification-warming impact was moderate, and it was mainly related to the pathways of metabolism, stress response, apoptosis, cell cycle, cell adhesion, and signaling for TFG-β and Hippo. The analysis of target mRNAs suggested that lncRNAs might contribute to the regulation of mRNAs after vitrification-warming. Our findings indicated that long-term storage after vitrification does not affect the mRNA and lncRNA expression profiles in human embryos, however, the procedure of vitrification-warming would lead to minor alteration of transcriptome.
Highlights
Since the first successful pregnancy from frozen embryos was reported in 1983, embryo cryopreservation has been widely used for over 30 years (Trounson and Mohr, 1983)
We aimed to explore the effect of the length of storage time on the mRNA and lncRNA expression profiles of vitrification cryopreserved human eight-cell embryos
To investigate the potential effects of storage time after vitrification on the transcriptomes of human embryos, we performed single-cell RNA-Seq on eleven donated human eight-cell embryos from three groups: fresh embryos, cryopreserved embryos stored for 3 years, and cryopreserved embryos stored for 8 years
Summary
Since the first successful pregnancy from frozen embryos was reported in 1983, embryo cryopreservation has been widely used for over 30 years (Trounson and Mohr, 1983). The application of cryopreservation technology allows multiple embryo transfers from a single stimulation cycle, which improves the cumulative live birth rate (Pandian et al, 2005; Zhu et al, 2018). Slow freezing and vitrification have been adopted as two principal cryopreservation methods (Edgar and Gook, 2012). Vitrification is a fast cryopreservation method that allows solidification of the cell(s) and extracellular milieu into a glass-like state, preventing the formation of ice crystals and cell damage (Rienzi et al, 2017). Several studies have demonstrated that vitrification can significantly increase the survival rate, clinical pregnancy rate, Effects of Long-Term Cryopreservation and live birth rate compared with slow freezing (AbdelHafez et al, 2010; Edgar and Gook, 2012). Owing to the improvement of clinical outcomes with vitrification, many laboratories from worldwide have completely replaced slow freezing with vitrification procedure
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