Effects of vitrification and cryostorage duration on single-cell RNA-Seq profiling of vitrified-thawed human metaphase II oocytes

Huo, Ying; Yuan, Peng; Qin, Qingyuan; Yan, Zhiqiang; Yan, Liying; Liu, Ping; Li, Rong; Yan, Jie

doi:10.1007/s11684-020-0792-7

Cited by 34 publications

(35 citation statements)

References 56 publications

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“…This caveat is somehow supported by evidence from two previous studies where it was shown that when these two proteins were knocked out, embryo development was blocked in 1-cell [28] and 2-cell embryos [25]. Similarly, previous reports have indicated that cryopreservation can lead to an overall decrease of mRNA extracted from cryopreserved MII oocytes compared to the control group [29,76]. Interestingly, a transcriptome-based analysis indicated that several downregulated genes were enriched in carbon metabolism, metabolic pathways, base excision repair, microtubule-based process, methylation, RNA transport, and cellular response to heat pathways and processes.…”

Section: Discussionmentioning

confidence: 75%

“…Interestingly, a transcriptome-based analysis indicated that several downregulated genes were enriched in carbon metabolism, metabolic pathways, base excision repair, microtubule-based process, methylation, RNA transport, and cellular response to heat pathways and processes. In particular, some of the downregulated genes in vitrified oocytes were also enriched in the MAPK signaling pathway [ 76 ]. Given that the MAPK signaling pathway plays an important role in regulating the maternal mRNA translation and degradation [ 71 ], therefore, we believe that the MAPK signaling pathway might regulate translation or degradation of those key maternal mRNAs to mitigate the negative effects induced by vitrification.…”

Section: Discussionmentioning

confidence: 99%

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Melatonin improves the first cleavage of parthenogenetic embryos from vitrified–warmed mouse oocytes potentially by promoting cell cycle progression

Pan

Qazi

Guo

et al. 2021

J Animal Sci Biotechnol

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Background This study investigated the effect of melatonin (MT) on cell cycle (G1/S/G2/M) of parthenogenetic zygotes developed from vitrified-warmed mouse metaphase II (MII) oocytes and elucidated the potential mechanism of MT action in the first cleavage of embryos. Results After vitrification and warming, oocytes were parthenogenetically activated (PA) and in vitro cultured (IVC). Then the spindle morphology and chromosome segregation in oocytes, the maternal mRNA levels of genes including Miss, Doc1r, Setd2 and Ythdf2 in activated oocytes, pronuclear formation, the S phase duration in zygotes, mitochondrial function at G1 phase, reactive oxygen species (ROS) level at S phase, DNA damage at G2 phase, early apoptosis in 2-cell embryos, cleavage and blastocyst formation rates were evaluated. The results indicated that the vitrification/warming procedures led to following perturbations 1) spindle abnormalities and chromosome misalignment, alteration of maternal mRNAs and delay in pronucleus formation, 2) decreased mitochondrial membrane potential (MMP) and lower adenosine triphosphate (ATP) levels, increased ROS production and DNA damage, G1/S and S/G2 phase transition delay, and delayed first cleavage, and 3) increased early apoptosis and lower levels of cleavage and blastocyst formation. Our results further revealed that such negative impacts of oocyte cryopreservation could be alleviated by supplementation of warming, recovery, PA and IVC media with 10− 9 mol/L MT before the embryos moved into the 2-cell stage of development. Conclusions MT might promote cell cycle progression via regulation of MMP, ATP, ROS and maternal mRNA levels, potentially increasing the first cleavage of parthenogenetic zygotes developed from vitrified–warmed mouse oocytes and their subsequent development.

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Section: Discussionmentioning

confidence: 75%

Section: Discussionmentioning

confidence: 99%

Melatonin improves the first cleavage of parthenogenetic embryos from vitrified–warmed mouse oocytes potentially by promoting cell cycle progression

Pan

Qazi

Guo

et al. 2021

J Animal Sci Biotechnol

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“…In agreement, recent data of single-cell RNA sequencing demonstrated that the vitrification procedure of human oocytes impaired their gene expression when compared with fresh oocytes. On the other hand, distinct periods of cryopreservation did not alter gene expression of oocytes submitted to vitrification [ 102 ]. Although these findings have contributed to understanding the impact of cryopreservation methods on germ cells, a deeper analysis of the transcriptomic changes at single-cell resolution of embryos and the effect of slow-freezing procedures on embryos and gametes is still needed.…”

Section: Molecular Alterations In Cryopreserved Oocytesmentioning

confidence: 99%

Cryopreservation of Gametes and Embryos and Their Molecular Changes

Estudillo

Jiménez

Bustamante-Nieves

et al. 2021

IJMS

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The process of freezing cells or tissues and depositing them in liquid nitrogen at –196 °C is called cryopreservation. Sub-zero temperature is not a physiological condition for cells and water ice crystals represent the main problem since they induce cell death, principally in large cells like oocytes, which have a meiotic spindle that degenerates during this process. Significantly, cryopreservation represents an option for fertility preservation in patients who develop gonadal failure for any condition and those who want to freeze their germ cells for later use. The possibility of freezing sperm, oocytes, and embryos has been available for a long time, and in 1983 the first birth with thawed oocytes was achieved. From the mid-2000s forward, the use of egg vitrification through intracytoplasmic sperm injection has improved pregnancy rates. Births using assisted reproductive technologies (ART) have some adverse conditions and events. These risks could be associated with ART procedures or related to infertility. Cryopreservation generates changes in the epigenome of gametes and embryos, given that ART occurs when the epigenome is most vulnerable. Furthermore, cryoprotective agents induce alterations in the integrity of germ cells and embryos. Notably, cryopreservation extensively affects cell viability, generates proteomic profile changes, compromises crucial cellular functions, and alters sperm motility. This technique has been widely employed since the 1980s and there is a lack of knowledge about molecular changes. The emerging view is that molecular changes are associated with cryopreservation, affecting metabolism, cytoarchitecture, calcium homeostasis, epigenetic state, and cell survival, which compromise the fertilization in ART.

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“…Data revealed that genes encoding proteins essential for oocyte development and specific functions (bone morphogenetic protein 15 (BMP15), growth differentiation factor 9 (GDF9), folliculogenesis-specific basic helix-loop-helix (FIGLA), POU Class 5 homeobox 1 (POU5f1-OCT4) and TATA box binding protein-associated factor 4B (AF4B)) were not altered by vitrification using a commercial vitrification kit [154]. The number of genes studied in 2021 was between fewer than 10 and almost 2000 [155]. Differentially expressed genes were found in vitrified-thawed oocytes compared to fresh MII oocytes (1646 and 341 genes were downregulated and upregulated, respectively) [155].…”

Section: Humansmentioning

confidence: 99%

“…The number of genes studied in 2021 was between fewer than 10 and almost 2000 [155]. Differentially expressed genes were found in vitrified-thawed oocytes compared to fresh MII oocytes (1646 and 341 genes were downregulated and upregulated, respectively) [155]. In cryopreserved oocytes, genes related to oxidative phosphorylation, the lysosome pathway, regulation of lipolysis in adipocytes or the AMPK signaling pathway were upregulated [155].…”

Section: Humansmentioning

confidence: 99%

Oocyte Cryopreservation in Domestic Animals and Humans: Principles, Techniques and Updated Outcomes

Tharasanit

Thuwanut

2021

Animals

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Oocyte cryopreservation plays important roles in basic research and the application of models for genetic preservation and in clinical situations. This technology provides long-term storage of gametes for genetic banking and subsequent use with other assisted reproductive technologies. Until recently, oocytes have remained the most difficult cell type to freeze, as the oocytes per se are large with limited surface area to cytoplasm ratio. They are also highly sensitive to damage during cryopreservation, and therefore the success rate of oocyte cryopreservation is generally poor when compared to noncryopreserved oocytes. Although advancement in oocyte cryopreservation has progressed rapidly for decades, the improvement of cryosurvival and clinical outcomes is still required. This review focuses on the principles, techniques, outcomes and prospects of oocyte cryopreservation in domestic animals and humans.

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Effects of vitrification and cryostorage duration on single-cell RNA-Seq profiling of vitrified-thawed human metaphase II oocytes

Cited by 34 publications

References 56 publications

Melatonin improves the first cleavage of parthenogenetic embryos from vitrified–warmed mouse oocytes potentially by promoting cell cycle progression

Melatonin improves the first cleavage of parthenogenetic embryos from vitrified–warmed mouse oocytes potentially by promoting cell cycle progression

Cryopreservation of Gametes and Embryos and Their Molecular Changes

Oocyte Cryopreservation in Domestic Animals and Humans: Principles, Techniques and Updated Outcomes

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