Site specificity of blastocyst hatching significantly influences pregnancy outcomes in mice

An, Liyou; Liu, Yanhong; Li, Mingyang; Liu, Zhihui; Wang, Zhisong; Dai, Yujian; Presicce, Giorgio Antonio; Du, Fuliang

doi:10.1096/fj.202100653r

Cited by 7 publications

(4 citation statements)

References 48 publications

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“…Quantitative RT‐PCR was performed using EvaGreen 2 × qPCR MasterMix‐ROX (Abm). Relative mRNA transcript levels were calculated using the

2^{- ∆ ∆ Ct}

method 22 . Gene expression levels of the GV group were normalized to 1.0 for comparison of expression between different stages of fertilized and PA embryos.…”

Section: Methodsmentioning

confidence: 99%

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Overcoming the H4K20me3 epigenetic barrier improves somatic cell nuclear transfer reprogramming efficiency in mice

et al. 2023

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Epigenetic reprogramming during fertilization and somatic cell nuclear transfer (NT) is required for cell plasticity and competent development. Here, we characterize the epigenetic modification pattern of H4K20me3, a repressive histone signature in heterochromatin, during fertilization and NT reprogramming. Importantly, the dynamic H4K20me3 signature identified during preimplantation development in fertilized embryos differed from NT and parthenogenetic activation (PA) embryos. In fertilized embryos, only maternal pronuclei carried the canonical H4K20me3 peripheral nucleolar ring‐like signature. H4K20me3 disappeared at the 2‐cell stage and reappeared in fertilized embryos at the 8‐cell stage and in NT and PA embryos at the 4‐cell stage. H4K20me3 intensity in 4‐cell, 8‐cell, and morula stages of fertilized embryos was significantly lower than in NT and PA embryos, suggesting aberrant regulation of H4K20me3 in PA and NT embryos. Indeed, RNA expression of the H4K20 methyltransferase Suv4‐20h2 in 4‐cell fertilized embryos was significantly lower than NT embryos. Knockdown of Suv4‐20h2 in NT embryos rescued the H4K20me3 pattern similar to fertilized embryos. Compared to control NT embryos, knockdown of Suv4‐20h2 in NT embryos improved blastocyst development ratios (11.1% vs. 30.5%) and full‐term cloning efficiencies (0.8% vs. 5.9%). Upregulation of reprogramming factors, including Kdm4b, Kdm4d, Kdm6a, and Kdm6b, as well as ZGA‐related factors, including Dux, Zscan4, and Hmgpi, was observed with Suv4‐20h2 knockdown in NT embryos. Collectively, these are the first findings to demonstrate that H4K20me3 is an epigenetic barrier of NT reprogramming and begin to unravel the epigenetic mechanisms of H4K20 trimethylation in cell plasticity during natural reproduction and NT reprogramming in mice.

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“…Quantitative RT‐PCR was performed using EvaGreen 2 × qPCR MasterMix‐ROX (Abm). Relative mRNA transcript levels were calculated using the

2^{- ∆ ∆ Ct}

method 22 . Gene expression levels of the GV group were normalized to 1.0 for comparison of expression between different stages of fertilized and PA embryos.…”

Section: Methodsmentioning

confidence: 99%

“…Relative mRNA transcript levels were calculated using the

method. 22 Gene expression levels of the GV group were normalized to 1.0 for comparison of expression between different stages of fertilized and PA embryos. Due to the limited number of NT embryos, RNA expression by RT‐PCR was not evaluated at different developmental stages.…”

Section: Methodsmentioning

confidence: 99%

Overcoming the H4K20me3 epigenetic barrier improves somatic cell nuclear transfer reprogramming efficiency in mice

et al. 2023

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“…A human expanded blastocyst (at day 5) created using a donor oocyte which underwent in vitro fertilization and culture has a diameter of around 194 µm [20]. For comparison, a murine expanded blastocyst (collected from a female mouse at day 3.5 post mating) has a diameter of 104 µm [21]. Embryonic genome activation (EGA) happens at different developmental stages and varies among species [22].…”

Section: Introductionmentioning

confidence: 99%

Development of a Microfluidic Chip Powered by EWOD for <em>In Vitro</em> Manipulation of Bovine Embryos

Karcz¹,

Soom²,

Smits³

et al. 2023

Preprint

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Digital microfluidics (DMF) holds great potential for the alleviation of laboratory procedures in assisted reproductive technologies (ARTs). Electrowetting on dielectric (EWOD) provides dynamic culture conditions in vitro that may better mimic the natural embryo microenvironment. Thus far, EWOD microdevices have been proposed for in vitro gamete and embryo handling in mice and for analyzing the human embryo secretome. This article presents the development of the first microfluidic chip utilizing EWOD technology designed for the manipulation of bovine embryos in vitro. The prototype sustains the cell cycles of embryos manipulated individually on the chips during in vitro culture (IVC). Challenges related to the chip fabrication as well as to its application during bovine embryo IVC in accordance with the adapted on-chip protocol are thoroughly discussed, and future directions for DMF in ARTs are indicated.

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“…A human expanded blastocyst (at day 5) created using a donor oocyte, which underwent in vitro fertilization and culture has a diameter of approximately 194 µm [ 24 ]. For comparison, a murine expanded blastocyst (collected from a female mouse at day 3.5 post mating) has a diameter of 104 µm [ 25 ]. Embryonic genome activation (EGA) happens at different developmental stages and varies among species [ 26 ].…”

Section: Introductionmentioning

confidence: 99%

Development of a Microfluidic Chip Powered by EWOD for In Vitro Manipulation of Bovine Embryos

et al. 2023

View full text Add to dashboard Cite

Digital microfluidics (DMF) holds great potential for the alleviation of laboratory procedures in assisted reproductive technologies (ARTs). The electrowetting on dielectric (EWOD) technology provides dynamic culture conditions in vitro that may better mimic the natural embryo microenvironment. Thus far, EWOD microdevices have been proposed for in vitro gamete and embryo handling in mice and for analyzing the human embryo secretome. This article presents the development of the first microfluidic chip utilizing EWOD technology designed for the manipulation of bovine embryos in vitro. The prototype sustains the cell cycles of embryos manipulated individually on the chips during in vitro culture (IVC). Challenges related to the chip fabrication as well as to its application during bovine embryo IVC in accordance with the adapted on-chip protocol are thoroughly discussed, and future directions for DMF in ARTs are indicated.

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Site specificity of blastocyst hatching significantly influences pregnancy outcomes in mice

Cited by 7 publications

References 48 publications

Overcoming the H4K20me3 epigenetic barrier improves somatic cell nuclear transfer reprogramming efficiency in mice

Overcoming the H4K20me3 epigenetic barrier improves somatic cell nuclear transfer reprogramming efficiency in mice

Development of a Microfluidic Chip Powered by EWOD for <em>In Vitro</em> Manipulation of Bovine Embryos

Development of a Microfluidic Chip Powered by EWOD for In Vitro Manipulation of Bovine Embryos

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