title: Histone modification of embryonic stem cells produced by somatic cell nuclear transfer and fertilized blastocysts.
abstract: OBJECTIVE
  Nuclear transfer-embryonic stem cells (NT-ESCs) are genetically identical to the donor's cells; provide a renewable source of tissue for replacement, and therefore, decrease the risk of immune rejection. Trichostatin A (TSA) as a histone deacetylase in- hibitor (HDACi) plays an important role in the reorganization of the genome and epigenetic changes. In this study, we examined whether TSA treatment after somatic cell nuclear transfer (SCNT) can improve the developmental rate of embryos and establishment rate of NT-ESCs line, as well as whether TSA treatment can improve histone modification in NT-ESCs lines.
  
  
  METHODS
  In this experimental study, mature oocytes were recovered from BDF1 [C57BL/6×DBA/2) F 1 mice] mice and enucleated by micromanipulator. Cumulus cells were injected into enucleated oocytes as donor. Reconstructed embryos were ac- tivated in the presence or absence of TSA and cultured for 5 days. Blastocysts were transferred on inactive mouse embryonic fibroblasts (MEF), so ESCs lines were estab- lished. ESCs markers were evaluated by reverse transcription-polymerase chain reaction (RT-PCR). Histone modifications were analyzed by enzyme linked immunosorbent assay (ELISA).
  
  
  RESULTS
  Result of this study showed that TSA treatment after SCNT can improve devel- opmental rate of embryos (21.12 ± 3.56 vs. 8.08 ± 7.92), as well as establishment rate of NT-ESCs line (25 vs. 12.5). We established 6 NT-ESCs in two experimental groups, and three embryonic stem cells (ESCs) lines as control group. TSA treatment has no effect in H3K4 acetylation and H3K9 tri-methylation in ESCs.
  
  
  CONCLUSIONS
  TSA plays a key role in the developmental rate of embryos, establishment rate of ESC lines after SCNT, and regulation of histone modification in NT-ESCs, in a man- ner similar to that of ESCs established from normal blastocysts.
authors.affiliation: 1.Cellular and Molecular Biology Research Center, Faculty of Medicine, Shahid Beheshti University of Medical Science, Tehran, Iran ; 2.Department of Cell Biology and Anatomical Science, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran ; 3.Department of Embryology, Mehr Infertility Institute, Rasht, Iran.
authors.forename: Fattaneh
authors.initials: F
authors.lastname: Farifteh
authors.affiliation: 1.Cellular and Molecular Biology Research Center, Faculty of Medicine, Shahid Beheshti University of Medical Science, Tehran, Iran ; 2.Department of Cell Biology and Anatomical Science, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
authors.forename: Mohammad
authors.initials: M
authors.lastname: Salehi
authors.affiliation: 1.Cellular and Molecular Biology Research Center, Faculty of Medicine, Shahid Beheshti University of Medical Science, Tehran, Iran ; 4.Department of Biothechnology, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
authors.forename: Mojgan
authors.initials: M
authors.lastname: Bandehpour
authors.affiliation: 5.Department of Immunology, Faculty of Medicine, Tehran, Iran.
authors.forename: Mosaffa
authors.initials: M
authors.lastname: Nariman
authors.affiliation: 2.Department of Cell Biology and Anatomical Science, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
authors.forename: Marefat
authors.initials: M
authors.lastname: Ghafari Novin
authors.affiliation: 6.Department of Molecular and Cellular Biology, University of Guelph, Canada.
authors.forename: Taher
authors.initials: T
authors.lastname: Hosseini
authors.affiliation: 7.Department of Transgenic Animal Science, Stem Cell Technology Research Center, Tehran, Iran.
authors.forename: Sedigheh
authors.initials: S
authors.lastname: Nematollahi
authors.affiliation: 2.Department of Cell Biology and Anatomical Science, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
authors.forename: Mohsen
authors.initials: M
authors.lastname: Noroozian
authors.affiliation: 3.Department of Embryology, Mehr Infertility Institute, Rasht, Iran.
authors.forename: Somayeh
authors.initials: S
authors.lastname: Keshavarzi
authors.affiliation: 1.Cellular and Molecular Biology Research Center, Faculty of Medicine, Shahid Beheshti University of Medical Science, Tehran, Iran ; 2.Department of Cell Biology and Anatomical Science, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran ; 3.Department of Embryology, Mehr Infertility Institute, Rasht, Iran.
authors.forename: Ahmad
authors.initials: A
authors.lastname: Hosseini
keywords: Epigenetics Modification; Somatic Cell Nuclear Transfer; Trichostatin A