Epigenetic states of donor cells significantly affect the development of somatic cell nuclear transfer (SCNT) embryos in pigs

Zhai, Yanhui; Li, Wei; Zhang, Zhiren; Cao, Yunqing; Wang, Zhengzhu; Zhang, Sheng; Li, Ziyi

doi:10.1002/mrd.22935

Cited by 31 publications

(33 citation statements)

References 44 publications

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“…However, AA had no effect on DNA methylation reprogramming in pig preimplantation embryos, because the antioxidant activity increases the blastocyst rate (Chawalit et al, ). Our results indicate that, similar to that of cattle (S. Zhang et al, ) and pigs (Zhai et al, ), the CDX2 promoter region is at a hypomethylation level during the yak IVF blastocyst stage. The difference was that the SOX2 , POU5F1 , and NANOG promoter regions are moderately methylated.…”

Section: Discussionsupporting

confidence: 84%

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DNA methylation regulated by ascorbic acids in yak preimplantation embryo helps to improve blastocyst quality

et al. 2019

Molecular Reproduction Devel

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DNA methylation as an important, essential epigenetic modification is critical for the successful development of mammalian embryos. In recent years, the important role of ascorbic acid (AA) as an irreplaceable cofactor for epigenetic regulation has been confirmed. However, the effect of AA on DNA methylation in preimplantation embryo development of plateau yak remains unknown. In this study, we explored whether AA can help regulates DNA methylation in yak preimplantation embryos to improve the blastocyst quality. First, our results indicate that the preimplantation of the yak still follows the classical pattern of DNA demethylation and remethylation, however, remethylation occurs in the blastocyst stage. Second, the unique expression pattern of the ten‐eleven translocation enzyme (TET3) in the cytoplasm plays a key role in the demethylation mechanism. Third, in the blastocyst stage, the pluripotency gene CDX2 promoter region was in a hypomethylated state, and the POU5F1, SOX2, and NANOG promoter regions were in moderate methylation states. In addition, treatment with 50 μg/ml AA mainly improved the expression levels of DNMT1, DNMT3a, and TET3, ensured the establishment, maintenance and transition of 5‐methylcytosine. After AA treatment, the methylation level of the pluripotency genes NANOG promoter regions was significantly reduced, and the mRNA transcript abundance of the pluripotency genes NANOG, POU5F1, and CDX2 was upregulated. In conclusion, our findings suggest that AA could increase blastocyst cell numbers by regulating DNA methylation of yak preimplantation embryos .

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Section: Discussionsupporting

confidence: 84%

“…pigs (Zhai et al, 2018), the CDX2 promoter region is at a hypomethylation level during the yak IVF blastocyst stage. The difference was that the SOX2, POU5F1, and NANOG promoter regions are moderately methylated.…”

Section: Discussionmentioning

confidence: 99%

DNA methylation regulated by ascorbic acids in yak preimplantation embryo helps to improve blastocyst quality

et al. 2019

Molecular Reproduction Devel

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“…High expression of OCT4 was more efficient as SCNT donor cells (Lee et al, ), which reinforces these results. Also, it has been well known that the epigenetic background can significantly affect embryonic development and the reprogramming efficiency of SCNT embryos (Zhai, Li, et al, ; Zhai, Zhang, et al, ). A representative example is a method in which a histone deacetylase inhibitors (HDACi) and DNA methyltransferase inhibitors (DNMTi) are treated with donor cells to improve cloning efficiency (Hou et al, ).…”

Section: Discussionmentioning

confidence: 99%

“…A representative example is a method in which a histone deacetylase inhibitors (HDACi) and DNA methyltransferase inhibitors (DNMTi) are treated with donor cells to improve cloning efficiency (Hou et al, ). According to Zhai, Li, et al (), the donor nuclei derived from bone marrow‐derived mesenchymal stem cells had more active epigenetic markers and fewer repressive epigenetic markers than FF. It could be considered that SSC had similar epigenetic characteristics of MSC, and this could influence nuclear reprogramming and improved blastocyst formation of SCNT embryos.…”

Section: Discussionmentioning

confidence: 99%

Comparative study of the developmental competence of cloned pig embryos derived from spermatogonial stem cells and fetal fibroblasts

Lee

et al. 2019

Reprod Domestic Animals

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Spermatogonial stem cells (SSC) are promising resources for genetic preservation and restoration of male germ cells in humans and animals. However, no studies have used SSC as donor nuclei in pig somatic cell nuclear transfer (SCNT). This study investigated the potential for use of porcine SSC as a nuclei donor for SCNT and developmental competence of SSC‐derived cloned embryos. In addition, demecolcine was investigated to determine whether it could prevent rupture of SSC during SCNT. When the potential of SSC to support embryonic development after SCNT was compared with that of foetal fibroblasts (FF), SSC‐derived SCNT embryos showed a higher (p < .05) developmental competence to the blastocyst stage (47.8%) than FF‐derived embryos (25.6%). However, when SSC were used as donor nuclei in the SCNT process, cell fusion rates were lower (p < .05) than when FF were used (61.9% vs. 75.8%). Treatment of SSC with demecolcine significantly (p < .05) decreased rupture of SSC during the SCNT procedure (7.5% vs. 18.8%) and increased fusion of cell‐oocyte couplets compared with no treatment (74.6% vs. 61.6%). In addition, SSC‐derived SCNT embryos showed higher blastocyst formation (48.4%) than FF‐derived embryos without (28.4%) and with demecolcine treatment (17.4%), even after demecolcine treatment. Our results demonstrate that porcine SSC are a desirable donor cell type for production of SCNT pig embryos and that demecolcine increases production efficiency of cloned embryos by inhibiting rupture of nuclei donor SSC during SCNT.

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“…However, when methylation level was lower in FIB compared to the MSC, as in BOV1, the use of TSA was not sufficient to increase the embryo production, since MSC were more methylated. According to Zhai et al (), the type and pattern of epigenetic modification in donor cells can significantly affect the developmental competency of SCNT embryos. Donor cells like MSCs, with a more‐open chromatin state, are more conducive to nuclear reprogramming, which improves nuclear transfer.…”

Section: Discussionmentioning

confidence: 99%

Use of trichostatin A alters the expression of HDAC3 and KAT2 and improves in vitro development of bovine embryos cloned using less methylated mesenchymal stem cells

Silva

Martins

Bessler

et al. 2018

Reprod Domestic Animals

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ContentsThe aim of this work was to investigate the methylation and hydroxymethylation status of mesenchymal stem cells (MSC) from amniotic fluid (MSC-AF), adipose tissue (MSC-AT) and fibroblasts (FIB-control) and to verify the effect of trichostatin A (TSA) on gene expression and development of cloned bovine embryos produced using these cells. Characterization of MSC from two animals (BOV1 and BOV2) was performed by flow cytometry, immunophenotyping and analysis of cellular differentiation genes expression. The cells were used in the nuclear transfer in the absence or presence of 50 nM TSA for 20 hr in embryo culture. Expression of HDAC1, HDAC3and KAT2A genes was measured in embryos by qRT-PCR. Methylation results showed difference between animals, with MSC from BOV2 demonstrating lower methylation rate than BOV1. Meanwhile, MSC-AF were less hydroxymethylated for both animals.MSC-AF from BOV2 produced 44.92 ± 8.88% of blastocysts when embryos were exposed to TSA and similar to embryo rate of MSC-AT also treated with TSA (37.96 ± 15.80%). However, when methylation was lower in FIB compared to MSC, as found in BOV1, the use of TSA was not sufficient to increase embryo production.MSC-AF embryos expressed less HDAC3 when treated with TSA, and expression of KAT2A was higher in embryos produced with all MSC and treated with TSA than embryos produced with FIB. The use of MSC less methylated and more hydroxymethylated in combination with embryo incubation with TSA can induce lower expression of HDAC3 and higher expression of KAT2A in the embryos and consequently improve bovine embryo production. K E Y W O R D Sadipose cells, amniotic cells, bovine cloning, DNA methylation, histone acetylation

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Epigenetic states of donor cells significantly affect the development of somatic cell nuclear transfer (SCNT) embryos in pigs

Cited by 31 publications

References 44 publications

DNA methylation regulated by ascorbic acids in yak preimplantation embryo helps to improve blastocyst quality

DNA methylation regulated by ascorbic acids in yak preimplantation embryo helps to improve blastocyst quality

Comparative study of the developmental competence of cloned pig embryos derived from spermatogonial stem cells and fetal fibroblasts

Use of trichostatin A alters the expression of HDAC3 and KAT2 and improves in vitro development of bovine embryos cloned using less methylated mesenchymal stem cells

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