Improvement of porcine cloning efficiency by trichostain A through early-stage induction of embryo apoptosis

Ji, Qiu; Zhu, Kangshun; Liu, Zhiguo; Song, Zhenwei; Huang, Yuankai; Zhao, Hongyong; Chen, Yaosheng; He, Zuyong; Mo, Delin; Peng, Cong

doi:10.1016/j.theriogenology.2012.12.010

Cited by 20 publications

(7 citation statements)

References 40 publications

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“…The oocytes were collected and cultured for 44 hr, as described in our previous study (Ji et al, ). Ovaries were collected from prepubertal gilts at a local abattoir, and transported to the laboratory in 0.9% (w/v) NaCl at 30–35°C within 2 hr.…”

Section: Methodsmentioning

confidence: 99%

“…The nuclear‐donor cells were porcine embryonic fibroblasts obtained from a Landrace fetus on Day 32 of pregnancy. Preparation of cell lines were carried out as previously described (Ji et al, ). The fibroblast suspension was centrifuged at 1,600 rpm for 5 min.…”

Section: Methodsmentioning

confidence: 99%

“…A terminal deoxynucleotidyl transferase‐mediated dUTP nick‐end labeling (TUNEL) assay was carried out as described previous (Ji et al, ). Blastocysts were fixed and permeabilized as above, washed three times in PBS–PVA, and incubated at 38.5°C in the dark for 30 min with TUNEL reaction medium (prepared immediately before use and kept on ice), covered with mineral oil.…”

Section: Methodsmentioning

confidence: 99%

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Exogenous expression of OCT4 facilitates oocyte‐mediated reprogramming in cloned porcine embryos

Peng

Zhao

et al. 2014

Molecular Reproduction Devel

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OCT4 is a well-established regulator of pluripotency and nuclear reprogramming. To determine if improving OCT4 abundance can facilitate oocyte-mediated reprogramming in cloned porcine embryos, we artificially increased OCT4 levels by co-incubating donor cells with 50 ng/µl OCT4 plasmid. We observed higher rates of blastocyst formation (P < 0.05) and lower levels of blastocyst apoptosis in nuclear-transfer-derived embryos carrying OCT4-incubated donor nuclei (OCT4-SCNT). The beneficial effect caused by exogenous expression of OCT4 involves epigenetic changes, wherein increased histone acetylation (AcH3K9) appeared in OCT4-SCNT embryos at the one-cell and blastocyst stages and reduced histone methylation (H3K9me2) was observed at the one-cell stage (P < 0.05). There was a transient increase in exogenous OCT4 and an up-regulation of endogenous OCT4 level in OCT4-SCNT embryos (P < 0.05), while the expression pattern of epigenetic enzymes was changed. These modifications were accompanied by an up-regulation of CDX2, whose interaction with OCT4 is instrumental for implantation, and a down-regulation of XIST, a negative indicator of reprogramming (P < 0.05). Taken together, our results support a role for exogenous expression of OCT4 in improving the efficiency of nuclear reprogramming while establishing a convenient and timesaving method to improve nuclear-transfer outcomes.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Exogenous expression of OCT4 facilitates oocyte‐mediated reprogramming in cloned porcine embryos

Peng

Zhao

et al. 2014

Molecular Reproduction Devel

Self Cite

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“…In this regard, one of the strategies for successful SCNT is reducing apoptosis to enhance the reprogramming efficiency by small molecules such as a histone deacetylase (HDAC) inhibitor, a ROS inhibitor and cytokines. For instance, a recent study revealed that trichostatin A (TSA), an inhibitor of HDAC, improved the developmental competence of cloned porcine embryos by improving epigenetic reprogramming [ 68 ]. The TSA-induced apoptosis during the initial embryonic stage allows abnormal cells to be eliminated, thus enhancing proliferation of healthy normal cells and improving quality.…”

Section: Reprogramming and Apoptosismentioning

confidence: 99%

Apoptosis in Porcine Pluripotent Cells: From ICM to iPSCs

Kim

Hyun

2016

IJMS

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Abstract:Pigs have great potential to provide preclinical models for human disease in translational research because of their similarities with humans. In this regard, porcine pluripotent cells, which are able to differentiate into cells of all three primary germ layers, might be a suitable animal model for further development of regenerative medicine. Here, we describe the current state of knowledge on apoptosis in pluripotent cells including inner cell mass (ICM), epiblast, embryonic stem cells (ESCs), and induced pluripotent stem cells (iPSCs). Information is focused on the apoptotic phenomenon in pluripotency, maintenance, and differentiation of pluripotent stem cells and reprogramming of somatic cells in pigs. Additionally, this review examines the multiple roles of apoptosis and summarizes recent progress in porcine pluripotent cells.

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“…Many variables contribute to improving efficiency (Ji et al. ); however, the donor cells were always recognized as an inconspicuous factor. Donor cells can reactivate their developmental plasticity in enucleated oocyte cytoplasm by nuclear transfer (NT), which contributes to totipotency in the artificially constructed embryos.…”

Section: Introductionmentioning

confidence: 99%

Establishment, Differentiation, Electroporation and Nuclear Transfer of Porcine Mesenchymal Stem Cells

Song

Peng

et al. 2015

Reprod Domestic Animals

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The limited success of somatic cell nuclear transfer (SCNT) is largely attributed to defects in epigenetic reprogramming of the donor genome. Donor cell types with distinct potential competence may offer different epigenetic flexibility for subsequent genome reprogramming in SCNT. Stem cells possibly enable their genomes to be more readily reprogrammed than differentiated cells. To improve the efficiency of cloning, porcine mesenchymal stem cells (pMSCs) were isolated and well identified by 6-channel flow cytometry and differentiation assays and were used as donors in SCNT. Compared with porcine embryonic fibroblasts (pEFs), our results showed that pMSCs markedly enhanced cloned embryo development in terms of cleavage and blastocyst formation (p < 0.05). To enhance the epigenetic flexibility of pMSCs, classical reprogramming factors (RFs) were transfected by electroporation, and we achieved optimization with ectopic expression of RFs in pMSCs. Our results suggest that the epigenetic status of donor cells has an improvement on genome reprogramming, and multipotent pMSCs favoured subsequent embryonic development.

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Improvement of porcine cloning efficiency by trichostain A through early-stage induction of embryo apoptosis

Cited by 20 publications

References 40 publications

Exogenous expression of OCT4 facilitates oocyte‐mediated reprogramming in cloned porcine embryos

Exogenous expression of OCT4 facilitates oocyte‐mediated reprogramming in cloned porcine embryos

Apoptosis in Porcine Pluripotent Cells: From ICM to iPSCs

Establishment, Differentiation, Electroporation and Nuclear Transfer of Porcine Mesenchymal Stem Cells

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