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논문검색

Effects of Trichostatin A on In Vitro Development of Porcine Parthenogenetic and Nuclear Transfer Embryos

초록

영어

Developmental potential of cloned embryos is related closely to epigenetic modification of somatic cell genome. The present study was to investigate the effects of applying histone deacetylation inhibitor, trichostatin A (TSA) to activated porcine embryos on subsequent development of porcine parthenogenetic and nuclear transfer embryos. Electrically activated oocytes were treated with 5 nM TSA for different exposure times (0, 1, 2 and 4 hr) and then the activated embryos were cultured for 7 days. The reconstructed embryos were treated with different concentrations of 0, 5, 10 and 25 nM TSA for 1 hr. Also 5 nM TSA was tested with different exposure times of 0, 0.5, 1, 2 and 4 hr. And fetal fibroblast cells were treated with 50 nM TSA for 1, 2 or 4 hr and with 5 nM TSA for 1 hr. Cumulus-free oocytes were enucleated and reconstructed by TSA-treated donor cells and electrically fused and cultured for 6 days. In parthenogenetic activation experiments, 5 nM TSA treatment for 1 hr significantly improved the percentage of blastocyst developmental rates than the other groups. Total cell number of blastocysts in 1 hr group was significantly higher than other groups or control. Similarly, blastocyst developmental rates of porcine NT embryos following 5 nM TSA treatment for 1 hr were highest. And the reconstructed embryos from donor cells treated by 50 nM TSA for 1 hr improved the percentage of blastocyst developmental rates than the control group. In conclusion, TSA treatment could improve the subsequent blastocyst development of porcine parthenogenetic and nuclear transfer embryos.

목차

ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
  Preparation of Porcine Fetal Fibroblasts
  Preparation of Oocytes
  Parthenogenetic Activation of Porcine Oocytes
  Nuclear Transfer and Fusion
  TSA Treatment
  Western Blot Analysis
  Experimental Design
  Statistical Analysis
 RESULTS
  Effects of TSA Exposure Times on In Vitro Parthenogenetic Pevelopment of Porcine Oocytes
  Effects of TSA Treatments on the Levels of Histone H3 Aceltylation of Fetal Fibroblast Cells
  Effects of TSA Treatments on the Development of PorcineNuclear Transfer Embryos Cultured In VitrEffects of TSA Treatments on the Development of Porcine Nuclear Transfer Embryos Cultured In Vitroo
 DISCUSSION
 REFERENCES

저자정보

  • Yun-Fei Diao Research Center for Transgenic and Cloned Pigs, Chungnam National University
  • Naruse Kenji Research Center for Transgenic and Cloned Pigs, Chungnam National University
  • Rong-Xun Han Research Center for Transgenic and Cloned Pigs, Chungnam National University
  • Tao-Lin Research Center for Transgenic and Cloned Pigs, Chungnam National University
  • Reza-K. Oqani Research Center for Transgenic and Cloned Pigs, Chungnam National University
  • Jung-Won Kang Research Center for Transgenic and Cloned Pigs, Chungnam National University
  • Dong-Il Jin Research Center for Transgenic and Cloned Pigs, Chungnam National University

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