5-Azacytidine improves the meiotic maturation and subsequent in vitro development of pig oocytes

Gupta, Mukesh Kumar; Heo, Young-Tae; Kim, Dong Ku; Lee, Hoon Taek; Uhm, Sang Jun

doi:10.1016/j.anireprosci.2019.106118

Cited by 16 publications

(6 citation statements)

References 51 publications

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“…A TSA-prompted decline in histone deacetylation may also affect other processes, such as the demethylation of DNA cytosine residues, incurring their intensification. A broad spectrum of non-specific HDACi, including TSA, and/or non-specific inhibitors of DNA methyltransferases (DNMTi) and/or selective inhibitors of histone methyltransferases (HMTi) have been formerly utilized in the strategies aimed to epigenomically modulate nuclear recipient oocytes [ 17 , 18 , 19 ], nuclear donor cells [ 20 , 21 , 22 , 23 ] and activated nuclear-transferred oocytes in pigs and other mammalian species [ 24 , 25 , 26 ]. These strategies have been developed to predominantly facilitate/refine the reprogramming of epigenomic memory and subsequently enhance the transcriptional activity of donor cell nuclear genomes in mammalian cloned embryos propagated by somatic cell nuclear transfer (SCNT) [ 27 , 28 , 29 , 30 , 31 ].…”

Section: Introductionmentioning

confidence: 99%

The Relative Abundances of Human Leukocyte Antigen-E, α-Galactosidase A and α-Gal Antigenic Determinants Are Biased by Trichostatin A-Dependent Epigenetic Transformation of Triple-Transgenic Pig-Derived Dermal Fibroblast Cells

Samiec

Wiater

Wartalski

et al. 2022

IJMS

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The present study sought to establish the mitotically stable adult cutaneous fibroblast cell (ACFC) lines stemming from hFUT2×hGLA×HLA-E triple-transgenic pigs followed by trichostatin A (TSA)-assisted epigenetically modulating the reprogrammability of the transgenes permanently incorporated into the host genome and subsequent comprehensive analysis of molecular signatures related to proteomically profiling the generated ACFC lines. The results of Western blot and immunofluorescence analyses have proved that the profiles of relative abundance (RA) noticed for both recombinant human α-galactosidase A (rhα-Gal A) and human leukocyte antigen-E (HLA-E) underwent significant upregulations in tri-transgenic (3×TG) ACFCs subjected to TSA-mediated epigenetic transformation as compared to not only their TSA-unexposed counterparts but also TSA-treated and untreated non-transgenic (nTG) cells. The RT-qPCR-based analysis of porcine tri-genetically engineered ACFCs revealed stable expression of mRNA fractions transcribed from hFUT2, hGLA and HLA-E transgenes as compared to a lack of such transcriptional activities in non-transgenic ACFC variants. Furthermore, although TSA-based epigenomic modulation has given rise to a remarkable increase in the expression levels of Galα1→3Gal (α-Gal) epitopes that have been determined by lectin blotting analysis, their semi-quantitative profiles have dwindled profoundly in both TSA-exposed and unexposed 3×TG ACFCs as compared to their nTG counterparts. In conclusion, thoroughly exploring proteomic signatures in such epigenetically modulated ex vivo models devised on hFUT2×hGLA×HLA-E triple-transgenic ACFCs that display augmented reprogrammability of translational activities of two mRNA transcripts coding for rhα-Gal A and HLA-E proteins might provide a completely novel and powerful research tool for the panel of further studies. The objective of these future studies should be to multiply the tri-transgenic pigs with the aid of somatic cell nuclear transfer (SCNT)-based cloning for the purposes of both xenografting the porcine cutaneous bioprostheses and dermoplasty-mediated surgical treatments in human patients.

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Section: Introductionmentioning

confidence: 99%

The Relative Abundances of Human Leukocyte Antigen-E, α-Galactosidase A and α-Gal Antigenic Determinants Are Biased by Trichostatin A-Dependent Epigenetic Transformation of Triple-Transgenic Pig-Derived Dermal Fibroblast Cells

Samiec

Wiater

Wartalski

et al. 2022

IJMS

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“…Many different determinants have been identified to be related to SCNT efficiency. They encompass (1) cytological and molecular quality of nuclear recipient oocytes characterized by meiotic, cytoplasmic, and epigenomic maturity parameters (Samiec & Skrzyszowska, 2012; Wang et al, 2018; Gupta et al, 2019); (2) the origin and phenotype of nuclear donor cells (Madheshiya et al, 2015; Em et al, 2016); (3) the capability of donor cell nuclear genome to be epigenetically reprogrammed in a cytoplasm of nuclear-transferred oocytes and descendant blastomeres of corresponding cloned embryos (Opiela et al, 2017; Monika & Selokar, 2018; Samiec & Skrzyszowska, 2018 a , 2018 b ; Zhou et al, 2020); (4) molecular and epigenetic communication between nuclear and mitochondrial genomes in nuclear-transferred oocytes and resultant cloned embryos (Hiendleder et al, 2003; Samiec, 2005 a , 2005 b ; Srirattana et al, 2011); (5) the strategies applied to artificially activate embryo-specific developmental programme of nuclear-transferred oocytes (Saikhun et al, 2004; Samiec & Skrzyszowska, 2010, 2014; George et al, 2011); and (6) the frequency of occurrence of apoptosis-dependent events in the in vitro -cultured nuclear donor cells and SCNT-derived embryos (Cui et al, 2011; Samiec et al, 2013; Agrawal et al, 2018; Mehta et al, 2019; Samiec et al, 2019).…”

Section: Introductionmentioning

confidence: 99%

Histone Demethylase KDM4D Could Improve the Developmental Competence of Buffalo (Bubalus Bubalis) Somatic Cell Nuclear Transfer (SCNT) Embryos

Feng

Zhao

et al. 2021

Microsc Microanal

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Somatic cell nuclear transfer (SCNT) holds vast potential in agriculture. However, its applications are still limited by its low efficiency. Histone 3 lysine 9 trimethylation (H3K9me3) was identified as an epigenetic barrier for this. Histone demethylase KDM4D could regulate the level of H3K9me3. However, its effects on buffalo SCNT embryos are still unclear. Thus, we performed this study to explore the effects and underlying mechanism of KDM4D on buffalo SCNT embryos. The results revealed that compared with the IVF embryos, the expression level of KDM4D in SCNT embryos was significantly lower at 8- and 16-cell stage, while the level of H3K9me3 in SCNT embryos was significantly higher at 2-cell, 8-cell, and blastocyst stage. Microinjection of KDM4D mRNA could promote the developmental ability of buffalo SCNT embryos. Furthermore, the expression level of ZGA-related genes such as ZSCAN5B, SNAI1, eIF-3a, and TRC at the 8-cell stage was significantly increased. Meanwhile, the pluripotency-related genes like POU5F1, SOX2, and NANOG were also significantly promoted at the blastocyst stage. The results were reversed after KDM4D was inhibited. Altogether, these results revealed that KDM4D could correct the H3K9me3 level, increase the expression level of ZGA and pluripotency-related genes, and finally, promote the developmental competence of buffalo SCNT embryos.

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“…Analogous enhancements in incidence of epigenetic reprogramming and improvements in transcriptional activities of genes in developing cloned embryos are affected by decreasing DNA methylation levels through exposure of in vitro cultured nuclear donor cells and/or nuclear recipient oocytes and/or SCNT-derived embryos to DNA methyltransferase inhibitors (DNMTi) (e.g., 5-aza-2'deoxycytidine (5-aza-dC)) (Enright et al, 2003;Gómez et al, 2012;. Finally, such HDACi-and/or DNMTi-dependent epigenomic modulation of nuclear donor cells, nuclear recipient cells or activated nuclear-transferred oocytes results in not only increasing in vitro and/or in vivo developmental capabilities of cloned embryos but also improving their cytological and molecular qualities (Samiec and Skrzyszowska, 2012b;Samiec et al, 2015Samiec et al, , 2019Opiela et al, 2017;Guo et al, 2018;Wang et al, 2018;Gupta et al, 2019). Histone methylation may also play an important role in somatic cell reprogramming.…”

Section: Molecular Aspects Of Somatic Cloningmentioning

confidence: 99%

Technical, Biological and Molecular Aspects of Somatic Cell Nuclear Transfer – A Review

Mrowiec

Bugno‐Poniewierska

2022

Annals of Animal Science

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Since the announcement of the birth of the first cloned mammal in 1997, Dolly the Sheep, 24 animal species including laboratory, farm, and wild animals have been cloned. The technique for somatic cloning involves transfer of the donor nucleus of a somatic cell into an enucleated oocyte at the metaphase II (MII) stage for the generation of a new individual, genetically identical to the somatic cell donor. There is increasing interest in animal cloning for different purposes such as rescue of endangered animals, replication of superior farm animals, production of genetically engineered animals, creation of biomedical models, and basic research. However, the efficiency of cloning remains relatively low. High abortion, embryonic, and fetal mortality rates are frequently observed. Moreover, aberrant developmental patterns during or after birth are reported. Researchers attribute these abnormal phenotypes mainly to incomplete nuclear remodeling, resulting in incomplete reprogramming. Nevertheless, multiple factors influence the success of each step of the somatic cloning process. Various strategies have been used to improve the efficiency of nuclear transfer and most of the phenotypically normal born clones can survive, grow, and reproduce. This paper will present some technical, biological, and molecular aspects of somatic cloning, along with remarkable achievements and current improvements.

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5-Azacytidine improves the meiotic maturation and subsequent in vitro development of pig oocytes

Cited by 16 publications

References 51 publications

The Relative Abundances of Human Leukocyte Antigen-E, α-Galactosidase A and α-Gal Antigenic Determinants Are Biased by Trichostatin A-Dependent Epigenetic Transformation of Triple-Transgenic Pig-Derived Dermal Fibroblast Cells

The Relative Abundances of Human Leukocyte Antigen-E, α-Galactosidase A and α-Gal Antigenic Determinants Are Biased by Trichostatin A-Dependent Epigenetic Transformation of Triple-Transgenic Pig-Derived Dermal Fibroblast Cells

Histone Demethylase KDM4D Could Improve the Developmental Competence of Buffalo (Bubalus Bubalis) Somatic Cell Nuclear Transfer (SCNT) Embryos

Technical, Biological and Molecular Aspects of Somatic Cell Nuclear Transfer – A Review

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