Porcine Granulocyte-Macrophage Colony-Stimulating Factor Improves the &lt;i&gt;in vitro&lt;/i&gt; Development of Cloned Porcine Embryos

Kwak, Seong-Sung; Cheong, Seung-A; Jeon, Yubyeol; Hyun, Sang‐Hwan

doi:10.1292/jvms.12-0050

Cited by 16 publications

(10 citation statements)

References 47 publications

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“…To further explore the effect of combined treatment-mediated the reduced 5mC and H3K9me2 on SCNT embryo development, we examined protein expression of two key transcription factors including OCT4 and CDX2 at the blastocyst stage after combined treatment or TSA treatment alone. The reduced expression of OCT4 and CDX2 in porcine somatic cloned blastocysts compared with IVF counterparts has been reported in the previous studies [26,35]. Analogous to this, we observed that expression levels of OCT4 and CDX2 in SCNT blastocysts were lower than that in in vivo and in vitro fertilized counterparts.…”

Section: Discussionsupporting

confidence: 89%

TSA and BIX-01294 Induced Normal DNA and Histone Methylation and Increased Protein Expression in Porcine Somatic Cell Nuclear Transfer Embryos

et al. 2017

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The poor efficiency of animal cloning is mainly attributed to the defects in epigenetic reprogramming of donor cells’ chromatins during early embryonic development. Previous studies indicated that inhibition of histone deacetylases or methyltransferase, such as G9A, using Trichostatin A (TSA) or BIX-01294 significantly enhanced the developmental efficiency of porcine somatic cell nuclear transfer (SCNT) embryos. However, potential mechanisms underlying the improved early developmental competence of SCNT embryos exposed to TSA and BIX-01294 are largely unclear. Here we found that 50 nM TSA or 1.0 μM BIX-01294 treatment alone for 24 h significantly elevated the blastocyst rate (P < 0.05), while further improvement was not observed under combined treatment condition. Furthermore, co-treatment or TSA treatment alone significantly reduced H3K9me2 level at the 4-cell stage, which is comparable with that in in vivo and in vitro fertilized counterparts. However, only co-treatment significantly decreased the levels of 5mC and H3K9me2 in trophectoderm lineage and subsequently increased the expression of OCT4 and CDX2 associated with ICM and TE lineage differentiation. Altogether, these results demonstrate that co-treatment of TSA and BIX-01294 enhances the early developmental competence of porcine SCNT embryos via improvements in epigenetic status and protein expression.

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Section: Discussionsupporting

confidence: 89%

TSA and BIX-01294 Induced Normal DNA and Histone Methylation and Increased Protein Expression in Porcine Somatic Cell Nuclear Transfer Embryos

et al. 2017

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“…In the mouse, blastocysts cultured with CSF2 in the medium experienced reduced expression of specific genes involved in stress responses and apoptosis (Chin et al, 2009) and fewer apoptotic cells after freeze/thawing (Desai et al 2007). In the pig, CSF2 affected gene expression in the blastocyst (Kwak et al 2012a) and increased the ratio of trophectoderm to inner cell mass (Kwak et al, 2012b). Embryos cultured with CSF2 have increased competence to develop to term after transfer to recipient females in the mouse (Sjöblom et al 2005), cow (Loureiro et al 2009; Denicol et al 2014b; embryos produced with X-sorted sperm), and human (Ziebe et al 2013).…”

Section: Introductionmentioning

confidence: 99%

Sex and the preimplantation embryo: implications of sexual dimorphism in the preimplantation period for maternal programming of embryonic development

et al. 2015

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The developmental program of the embryo displays a plasticity that can result in long-acting effects that extend into post-natal life. In mammals, adult phenotype can be altered by changes in the maternal environment during the preimplantation period. One characteristic of developmental programming during this time is that the change in adult phenotype is often different for female offspring than for male offspring. In this paper, we propose the hypothesis that sexual dimorphism in preimplantation programming is mediated, at least in part, by sex-specific responses of embryos to maternal regulatory molecules whose secretion is dependent on maternal environment. The strongest evidence for this idea comes from the study of colony-stimulating factor 2 (CSF2). Expression of CSF2 from the oviduct and endometrium is modified by environmental factors of the mother, in particular seminal plasma and obesity. Additionally, CSF2 alters several properties of the preimplantation embryo and has been shown to alleviate negative consequences of culture of mouse embryos on postnatal phenotype in a sex-dependent manner. In cattle, exposure of preimplantation bovine embryos to CSF2 causes sex-specific changes in gene expression, interferon-τ secretion, and DNA methylation later in pregnancy (day 15 of gestation). It is likely that several embryokines can alter postnatal phenotype through actions directed towards the preimplantation embryo. Identification of these molecules and elucidation of the mechanisms by which sexually-disparate programming is established will lead to new insights into the control and manipulation of embryonic development.

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“…Introducing mouse GM‐CSF into embryo culture increased in vitro development of parthenotes in a protein free culture condition (Cui et al, 2004). Pig GM‐CSF had a positive effect on the in vitro development of IVF‐ and SCNT‐derived embryos (Kwak et al, 2012a, b). While positive effects of GM‐CSF on embryos has been reported, all results are restricted only to in vitro studies, thus limiting its application in vivo.…”

Section: Introductionmentioning

confidence: 99%

Piglets produced from cloned blastocysts cultured in vitro with GM‐CSF

Lee

Redel

Spate

et al. 2013

Molecular Reproduction Devel

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SUMMARY In general, pig embryos established by somatic cell nuclear transfer (SCNT) are transferred at the one-cell stage because of suboptimal embryo culture conditions. Improvements in embryo culture can increase the practical application of late embryo transfer. The goal of this study was to evaluate embryos cultured with granulocyte-macrophage colony-stimulating factor (GM-CSF) in vitro, and to track the in vivo developmental competency of SCNT-derived blastocysts from these GM-CSF embryos. The receptor for GM-CSF was up-regulated in in vitro-produced embryos when compared to in vivo-produced cohorts, but the level decreased when GM-CSF was present. In vitro fertilized (IVF) embryos, supplemented with GM-CSF (2 or 10 ng/ml), showed a higher frequency of development to the blastocyst stage compared to controls. The total cell numbers of the blastocysts also increased with supplementation of GM-CSF. Molecular analysis demonstrates that IVF-derived blastocysts cultured with GM-CSF exhibit less apoptotic activity. Similarly, an increase in development to the blastocyst stage and an increase in the average total-cell number in the blastocysts were observed when SCNT-derived embryos were cultured with either concentration of GM-CSF (2 or 10 ng/ml). When SCNT-derived embryos, cultured with 10 ng/ml GM-CSF, were transferred into six surrogates at Day 6, five of the surrogates became pregnant and delivered healthy piglets. Our findings suggest that supplementation of GM-CSF can provide better culture conditions for IVF- and SCNT-derived embryos, and pig SCNT-derived embryos cultured with GM-CSF in vitro can successfully produce piglets when transferred into surrogates at the blastocyst stage. Thus, it may be practical to begin performing SCNT-derived embryo transfer at the blastocyst stage.

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Porcine Granulocyte-Macrophage Colony-Stimulating Factor Improves the <i>in vitro</i> Development of Cloned Porcine Embryos

Cited by 16 publications

References 47 publications

TSA and BIX-01294 Induced Normal DNA and Histone Methylation and Increased Protein Expression in Porcine Somatic Cell Nuclear Transfer Embryos

TSA and BIX-01294 Induced Normal DNA and Histone Methylation and Increased Protein Expression in Porcine Somatic Cell Nuclear Transfer Embryos

Sex and the preimplantation embryo: implications of sexual dimorphism in the preimplantation period for maternal programming of embryonic development

Piglets produced from cloned blastocysts cultured in vitro with GM‐CSF

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