Treatment of Donor Cells and Reconstructed Embryos with a Combination of Trichostatin-A and 5-aza-2'-Deoxycytidine Improves the Developmental Competence and Quality of Buffalo Embryos Produced by Handmade Cloning and Alters Their Epigenetic Status and Gen


:The application of cloning technology on a large scale is limited by very low offspring rate primarily due to aberrant or incomplete epigenetic reprogramming. Trichostatin A (TSA), a histone deacetylase inhibitor, and 5-aza-2'-deoxycytidine (5-aza-dC), an inhibitor of DNA methyltransferases, are widely used for altering the epigenetic status of cloned embryos. We optimized the doses of these epigenetic modifiers for production of buffalo embryos by handmade cloning and examined whether combined treatment with these epigenetic modifiers offered any advantage over treatment with the individual epigenetic modifier. Irrespective of whether donor cells or reconstructed embryos or both were treated with 50 nM TSA +7.5 nM 5-aza-dC, (1) the blastocyst rate was significantly higher (71.6 ± 3.5, 68.3 ± 2.6, and 71.8 ± 2.4, respectively, vs. 43.1 ± 3.4 for controls, p < 0.05); (2) the apoptotic index was lower (5.4 ± 1.1, 9.5 ± 1.0, and 7.4 ± 1.3, respectively, vs. 19.5 ± 2.1 for controls, p < 0.05) and was similar to that of in vitro fertilization blastocysts (6.0 ± 0.8); (3) the global level of H3K18ac was higher (p < 0.01) and that of H3K27me3 lower (p < 0.05) than in controls and was similar among all treatment groups; and (4) the expression level of epigenetic-(HDAC1, DNMT1, and DNMT3a), pluripotency-(OCT4 and NANOG), and development-related (FGF4) genes, but not that of SOX2 and CDX2, was similar among all treatment groups. These results demonstrate that similar levels of beneficial effects can be obtained following treatment of either donor cells or reconstructed embryos or both with the combination of TSA +5-aza-dC. Therefore, there is no advantage in treating both donor cells and reconstructed embryos when the combination of TSA and 5-aza-dC is used.


Cell Reprogram


Cellular reprogramming


Saini M,Selokar NL,Agrawal H,Singla SK,Chauhan MS,Manik RS,Palta P




Has Abstract


2017-06-01 00:00:00












  • Cellular transitions and tissue engineering.

    abstract::Epithelial-to-mesenchymal transition (EMT) and endothelial-to-mesenchymal transition (EndMT) describe complex changes in progenitor lineage, cell morphology, and gene expression. Stimulated by environmental cues, these cellular transitions are essential for elements of embryonic development and can be pathologically d...

    journal_title:Cellular reprogramming

    pub_type: 杂志文章,评审


    authors: Schindeler A,Kolind M,Little DG

    更新日期:2013-04-01 00:00:00

  • Expression of innate immune response genes in liver and three types of adipose tissue in cloned pigs.

    abstract::The pig has been proposed as a relevant model for human obesity-induced inflammation, and cloning may improve the applicability of this model. We tested the assumptions that cloning would reduce interindividual variation in gene expression of innate immune factors and that their expression would remain unaffected by t...

    journal_title:Cellular reprogramming

    pub_type: 杂志文章


    authors: Rødgaard T,Skovgaard K,Stagsted J,Heegaard PM

    更新日期:2012-10-01 00:00:00

  • Germ-line-competent embryonic stem cells of the Chinese Kunming mouse strain with long-term self-renewal ability.

    abstract::Kunming (KM) mice are the most widely used strain in China. However, authentic embryonic stem cells (ESCs) from KM mice have never been available, and this hampers the genetic manipulation of this valuable mice strain. In this study, we show that KM ESCs can be efficiently derived and maintained in chemically defined ...

    journal_title:Cellular reprogramming

    pub_type: 杂志文章


    authors: Peng X,Liu T,Yang B,Shi C,Sun Y,Jiang L,Jin H,Li L,Zhu H,Wu M,Qian Q

    更新日期:2013-06-01 00:00:00

  • Overexpression of MBD3 Improves Reprogramming of Cloned Pig Embryos.

    abstract::Methyl-CpG-binding domain protein 3 (MBD3) is a core component of the nucleosome remodeling and deacetylase (NuRD) complex, which is crucial for pluripotent stem cell differentiation and embryonic development. MBD3 was shown to play important roles in transcription factor-induced somatic cell reprogramming. Expression...

    journal_title:Cellular reprogramming

    pub_type: 杂志文章


    authors: Wang X,Shi J,Cai G,Zheng E,Liu D,Wu Z,Li Z

    更新日期:2019-10-01 00:00:00

  • Culture conditions and enzymatic passaging of bovine ESC-like cells.

    abstract::The goals of the current study were to (1) improve culture conditions and (2) chemical passaging of bovine embryonic stem cell-like (bESC-like) cells. Specifically, the effects of human leukemia inhibitory factor (hLIF), two types of feeders, mouse embryonic fibroblast (MEF) and bovine embryonic fibroblast (BEF), as w...

    journal_title:Cellular reprogramming

    pub_type: 杂志文章


    authors: Gong G,Roach ML,Jiang L,Yang X,Tian XC

    更新日期:2010-04-01 00:00:00

  • Kinetic analysis of porcine fibroblast reprogramming toward pluripotency by defined factors.

    abstract::Induced pluripotent stem cells (iPSCs) are generated from somatic cells through ectopic expression of defined transcription factors. So far, many iPSC lines have been established in various species, including porcine. However, the molecular events during somatic cell reprogramming in pig are largely unknown. The aim o...

    journal_title:Cellular reprogramming

    pub_type: 杂志文章


    authors: Cheng D,Li Z,Liu Y,Gao Y,Wang H

    更新日期:2012-08-01 00:00:00

  • Keith's MAGIC: Cloning and the Cell Cycle.

    abstract::Abstract Professor Keith Campbell's critical contribution to the discovery that a somatic cell from an adult animal can be fully reprogrammed by oocyte factors to form a cloned individual following nuclear transfer (NT)(Wilmut et al., 1997 ) overturned a dogma concerning the reversibility of cell fate that many scient...

    journal_title:Cellular reprogramming

    pub_type: 传,历史文章,杂志文章


    authors: Wells DN

    更新日期:2013-10-01 00:00:00

  • Hematopoietic Developmental Potential of Human Pluripotent Stem Cell Lines Is Accompanied by the Morphology of Embryoid Bodies and the Expression of Endodermal and Hematopoietic Markers.

    abstract::The potential clinical applications of hematopoietic stem cells (HSCs) derived from human pluripotent stem cells (hPSCs) are limited by the difficulty of recapitulating embryoid hematopoiesis and by the unknown differentiation potential of hPSC lines. To evaluate their hematopoietic developmental potential, available ...

    journal_title:Cellular reprogramming

    pub_type: 杂志文章


    authors: Tesarova L,Simara P,Stejskal S,Koutna I

    更新日期:2017-08-01 00:00:00

  • In Vitro Induction of Human Embryonic Stem Cells into the Midbrain Dopaminergic Neurons and Transplantation in Cynomolgus Monkey.

    abstract::A simple, rapid, efficient, and specialized culture system was successfully developed in this study to induce human embryonic stem cells into dopaminergic neurons in vitro. It only took 5 days to generate quickly and directly a large number of homogeneous neural stem cell (NSC) spheres by the introduction of small mol...

    journal_title:Cellular reprogramming

    pub_type: 杂志文章


    authors: Qiu X,Liu Y,Xiao X,He J,Zhang H,Li Y

    更新日期:2019-12-01 00:00:00

  • Retracted: Bone Mesenchymal Stem Cell-Conditioned Medium Regulates the Differentiation of Neural Stem Cells Via Notch Pathway Activation.

    abstract::The online-ahead-of print e-pub version of the article entitled, Bone Mesenchymal Stem Cell-Conditioned Medium Regulates the Differentiation of Neural Stem Cells Via Notch Pathway Activation, by Li H-M, Tong Y, Xia X, Huang J, Song P-W, Zhang R-J, Shen C-L, utilizing the DOI number 10.1089/cell.2018.0042 is being offi...

    journal_title:Cellular reprogramming

    pub_type: 杂志文章,收录出版


    authors: Li HM,Tong Y,Xia X,Huang J,Song PW,Zhang RJ,Shen CL

    更新日期:2018-12-27 00:00:00

  • Taillessness in a Cloned Cow is Not Genetically Transmitted.

    abstract::Somatic cell nuclear transfer (SCNT), commonly referred to as cloning, results in the generation of offspring that, except for mitochondrial DNA, are genetically identical to the nuclear donor. We previously used a genetically modified bovine cell line as the donor for SCNT and obtained a calf, named Daisy, that was b...

    journal_title:Cellular reprogramming

    pub_type: 杂志文章


    authors: Wagner S,Cullum A,Wells DN,Laible G

    更新日期:2017-12-01 00:00:00

  • Effects of histone acetylation status on the early development of in vitro porcine transgenic cloned embryos.

    abstract::The purpose of this study was to investigate the effects of the histone deacetylase (HDAC) inhibitor trichostatin A (TSA) on transgene expression and development of porcine transgenic cloned embryos, specifically focusing on effects derived from TSA-treated donor cells or TSA-treated reconstructed embryos. The results...

    journal_title:Cellular reprogramming

    pub_type: 杂志文章


    authors: Luo B,Ju S,Muneri CW,Rui R

    更新日期:2015-02-01 00:00:00

  • Gnotobiotic Miniature Pig Interbreed Somatic Cell Nuclear Transfer for Xenotransplantation.

    abstract::Transgenic animal producing technology has improved consistently over the last couple of decades. Among the available methods, somatic cell nuclear transfer (SCNT) technology was officially the most popular. However, SCNT has low efficiency and requires a highly skilled individual. Additionally, the allo-SCNT nuclear ...

    journal_title:Cellular reprogramming

    pub_type: 杂志文章


    authors: Hwang JH,Kim SE,Gupta MK,Lee H

    更新日期:2016-08-01 00:00:00

  • Improved production of genetically modified fetuses with homogeneous transgene expression after transgene integration site analysis and recloning in cattle.

    abstract::Animal cloning by nuclear transfer (NT) has made the production of transgenic animals using genetically modified donor cells possible and ensures the presence of the gene construct in the offspring. The identification of transgene insertion sites in donor cells before cloning may avoid the production of animals that c...

    journal_title:Cellular reprogramming

    pub_type: 杂志文章


    authors: Bressan FF,Dos Santos Miranda M,Perecin F,De Bem TH,Pereira FT,Russo-Carbolante EM,Alves D,Strauss B,Bajgelman M,Krieger JE,Binelli M,Meirelles FV

    更新日期:2011-02-01 00:00:00

  • Enucleation of demecolcine-treated bovine oocytes in cytochalasin-free medium: mechanism investigation and practical improvement.

    abstract::Demecolcine-assisted/induced enucleation has been used in nuclear transfer cloning procedures for many species, yet its mechanism of action remains unclear. Primarily because oocytoplasm protrusion induced by demecolcine is inhibited by the presence of cytochalasin, its use has had limited application. In this experim...

    journal_title:Cellular reprogramming

    pub_type: 杂志文章


    authors: Meng Q,Wu X,Bunch TD,White K,Sessions BR,Davies CJ,Rickords L,Li GP

    更新日期:2011-10-01 00:00:00

  • Cloned sheep blastocysts derived from oocytes enucleated manually using a pulled pasteur pipette.

    abstract::The potential applications of a simplified method of somatic cell nuclear transfer (SCNT) that is improved in both efficiency and throughput is considerable. Technically, a major step of SCNT is to produce large pools of enucleated oocytes (cytoplasts) efficiently, a process that requires considerable micromanipulatio...

    journal_title:Cellular reprogramming

    pub_type: 杂志文章


    authors: Hosseini SM,Hajian M,Moulavi F,Asgari V,Forouzanfar M,Nasr-Esfahani MH

    更新日期:2013-02-01 00:00:00

  • Nonviral minicircle generation of induced pluripotent stem cells compatible with production of chimeric chickens.

    abstract::Chickens are vitally important in numerous countries as a primary food source and a major component of economic development. Efforts have been made to produce transgenic birds through pluripotent stem cell [primordial germ cells and embryonic stem cells (ESCs)] approaches to create animals with improved traits, such a...

    journal_title:Cellular reprogramming

    pub_type: 杂志文章


    authors: Yu P,Lu Y,Jordan BJ,Liu Y,Yang JY,Hutcheson JM,Ethridge CL,Mumaw JL,Kinder HA,Beckstead RB,Stice SL,West FD

    更新日期:2014-10-01 00:00:00

  • Murine embryonic fibroblast cell lines differentiate into three mesenchymal lineages to different extents: new models to investigate differentiation processes.

    abstract::Various diseases, injuries, and congenital abnormalities may result in degeneration and loss of organs and tissues. Recently, tissue engineering has offered new treatment options for these common, severe, and costly problems in human health care. Its application is often based on the usage of differentiated stem cells...

    journal_title:Cellular reprogramming

    pub_type: 杂志文章


    authors: Dastagir K,Reimers K,Lazaridis A,Jahn S,Maurer V,Strauß S,Dastagir N,Radtke C,Kampmann A,Bucan V,Vogt PM

    更新日期:2014-08-01 00:00:00

  • Development and gene expression of porcine cloned embryos derived from bone marrow stem cells with overexpressing Oct4 and Sox2.

    abstract::The present study compared the potential of porcine bone marrow mesenchymal stem cells (pBMSCs) at different passages as nuclear transfer (NT) donors and the developmental efficiency of NT embryos from donor cells transfected with/without Oct4 and Sox2. Early-passage pBMSCs showed higher proliferation and expression o...

    journal_title:Cellular reprogramming

    pub_type: 杂志文章


    authors: Lee JH,Lee WJ,Jeon RH,Lee YM,Jang SJ,Lee SL,Jeon BG,Ock SA,King WA,Rho GJ

    更新日期:2014-12-01 00:00:00

  • Effect of the time interval between fusion and activation on epigenetic reprogramming and development of bovine somatic cell nuclear transfer embryos.

    abstract::Previous studies have shown that the time interval between fusion and activation (FA interval) play an important role in nuclear remodeling and in vitro development of somatic cell nuclear transfer (SCNT) embryos. However, the effects of FA interval on the epigenetic reprogramming and in vivo developmental competence ...

    journal_title:Cellular reprogramming

    pub_type: 杂志文章


    authors: Liu J,Wang Y,Su J,Wang L,Li R,Li Q,Wu Y,Hua S,Quan F,Guo Z,Zhang Y

    更新日期:2013-04-01 00:00:00

  • Posttranslational modifications of defined embryonic reprogramming transcription factors.

    abstract::The generation of induced pluripotent stem cells (iPSCs) from somatic cells by expressing ectopic reprogramming transcriptional factors such as Oct3/4, Sox2, Klf4, c-Myc, and Nanog is one of the cutting-edge discoveries in stem cell and cancer research. This discovery has raised several safety issues regarding the use...

    journal_title:Cellular reprogramming

    pub_type: 杂志文章,评审


    authors: Ramakrishna S,Kim KS,Baek KH

    更新日期:2014-04-01 00:00:00

  • Selective isolation of nanog-positive human amniotic mesenchymal cells and differentiation into cardiomyocytes.

    abstract::Adult cardiomyocytes have little ability to regenerate, thus cardiac regeneration therapy represents a potential method for treating severe heart failure. Human amniotic mesenchymal cells (hAMCs) have the potential to be a useful cell source for cardiac regeneration therapy. We attempted to isolate stem cells from hAM...

    journal_title:Cellular reprogramming

    pub_type: 杂志文章


    authors: Otaka S,Nagura S,Koike C,Okabe M,Yoshida T,Fathy M,Yanagi K,Misaki T,Nikaido T

    更新日期:2013-02-01 00:00:00

  • Acclimatized induction reveals the multipotency of adult human undifferentiated keratinocytes.

    abstract::It has been demonstrated that several types of somatic stem cells have the remarkable capacity to differentiate into other types of tissues. However, the promise of keratinocyte stem cells seems slim for generating nonepidermal tissues. Using our recently developed acclimatization induction strategy, we demonstrate th...

    journal_title:Cellular reprogramming

    pub_type: 杂志文章


    authors: Han W,Chen M,Li M,Wu Z,Zhao Y,Wang Y,Wang L,Yu L,Fu X

    更新日期:2010-06-01 00:00:00

  • Improved bovine ICSI outcomes by sperm selected after combined heparin-glutathione treatment.

    abstract::Despite widespread application of intracytoplasmic sperm injection (ICSI) in human-assisted reproductive techniques (ART), the efficiency of this method is still far from satisfactory in livestock, particularly in the bovine species with its unique sperm condensation. On the basis of the natural chemical structure of ...

    journal_title:Cellular reprogramming

    pub_type: 杂志文章


    authors: Sekhavati MH,Shadanloo F,Hosseini MS,Tahmoorespur M,Nasiri MR,Hajian M,Nasr-Esfahani MH

    更新日期:2012-08-01 00:00:00

  • Generation of induced pluripotent stem cells from hair follicle bulge neural crest stem cells.

    abstract::Induced pluripotent stem cells (iPSCs) are promising candidates for the study of disease models as well as for tissue engineering purposes. Part of a strategy to develop safe reprogramming technique is reducing the number of exogenous reprogramming factors. Some cells types are more prone to reprogramming than others....

    journal_title:Cellular reprogramming

    pub_type: 杂志文章


    authors: Ma MS,Czepiel M,Krause T,Schäfer KH,Boddeke E,Copray S

    更新日期:2014-10-01 00:00:00

  • Ovine oocytes vitrified at germinal vesicle stage as cytoplast recipients for somatic cell nuclear transfer (SCNT).

    abstract::The development of embryos produced by somatic cell nuclear transfer (SCNT) using vitrified oocytes as cytoplast recipients has been reported in cattle but not in sheep. This study investigated the parthenogenetic development of ovine oocytes vitrified and thawed at the germinal vesicle (GV) stage, matured in vitro, a...

    journal_title:Cellular reprogramming

    pub_type: 杂志文章


    authors: Moawad AR,Choi I,Zhu J,Campbell KH

    更新日期:2011-08-01 00:00:00

  • Factors Affecting Mouse Somatic Cell Nuclear Reprogramming by Rabbit Ooplasms.

    abstract::Successful development of interspecies somatic cell nuclear transfer (iSCNT) embryos depends on compatibilities between ooplasmic and nuclear components. However, the mechanisms by which the compatibilities are regulated are still unknown. In this study, using mouse Oct4-green fluorescent protein (GFP) cells as donors...

    journal_title:Cellular reprogramming

    pub_type: 杂志文章


    authors: Huang X,Song L,Zhan Z,Gu H,Feng H,Li Y

    更新日期:2017-12-01 00:00:00

  • Effect of POU5F1 Expression Level in Clonal Subpopulations of Bovine Fibroblasts Used as Nuclear Donors for Somatic Cell Nuclear Transfer.

    abstract::Somatic cell nuclear transfer (SCNT) success is partially hindered by the low epigenetic reprogramming efficiency of the donor cell. Previous studies suggest cellular heterogeneity among donor nuclei in regard to reprogramming potential, which precludes comparison among different strategies to increase cloning success...

    journal_title:Cellular reprogramming

    pub_type: 杂志文章


    authors: Sá AL,Sampaio RV,da Costa Almeida NN,Sangalli JR,Brito KNL,Bressan FF,Rissino JD,do Socorro Damasceno Santos S,Meirelles FV,Ohashi OM,Dos Santos Miranda M

    更新日期:2017-10-01 00:00:00

  • Sheep: the first large animal model in nuclear transfer research.

    abstract::The scope of this article is not to provide an exhaustive review of nuclear transfer research, because many authoritative reviews exist on the biological issues related to somatic and embryonic cell nuclear transfer. We shall instead provide an overview on the work done specifically on sheep and the value of this work...

    journal_title:Cellular reprogramming

    pub_type: 杂志文章,评审


    authors: Loi P,Czernik M,Zacchini F,Iuso D,Scapolo PA,Ptak G

    更新日期:2013-10-01 00:00:00

  • Hypoxia Enhances Direct Reprogramming of Mouse Fibroblasts to Cardiomyocyte-Like Cells.

    abstract::Recent work has shown that mouse and human fibroblasts can be reprogrammed to cardiomyocyte-like cells with a combination of transcription factors. Current research has focused on improving the efficiency and mechanisms for fibroblast reprogramming. Previously, it has been reported that hypoxia enhances fibroblast cel...

    journal_title:Cellular reprogramming

    pub_type: 杂志文章


    authors: Wang Y,Shi S,Liu H,Meng L

    更新日期:2016-02-01 00:00:00