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논문검색

Effect of Osmolarity of Culture Medium on Imprinting and Apoptotic Gene Expression in Miniature Pig Nuclear Transfer Embryos

초록

영어

This study was conducted to investigate the development and gene expression in miniature pig nuclear transfer (mNT) embryos produced under different osmolarity culture conditions. Control group of mNT embryos was cultured in PZM-3 for 6 days. Treatment group of mNT embryos was cultured in modified PZM-3 with NaCl (mPZM-3, 320 mOsmol) for 2 days, and then cultured in PZM-3 (270 mOsmol) for 4 days. Blastocyst formation rate of the treatment group was significantly higher than the control and the apoptosis rate was significantly lower in treatment group. Bax- and caspase-3 mRNA expression were significantly higher in the control than the treatment group. Also, the majority of imprinting genes were expressed aberrantly in in vitro produced mNT blastocysts compared to in vivo derived blastocyst H19 and Xist mRNA expression were significantly lower in the control than the treatment group or in vivo. IGF2 mRNA expression was significantly higher in the control than the treatment group or in vivo. IGF2r mRNA expression was significantly lower in the control. Methylation profiles of individual DNA strands in H19 upstream T-DMR sequences showed a similar methylation status between treatment group and in vivo. These results indicate that the modification of osmolarity in culture medium at early culture stage could provide more beneficial culture environments for mNT embryos.

목차

ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
  Oocyte Recovery and Maturation
  Preparation of Nuclear Donor Cells
  Production of Nuclear Transfer Embryos
  Terminal DeoxynucleotidyI Transferase-Mediated dUTP Nick-End Labeling (TUNEL) Assay
  Collection of In Vivo Blastocysts
  Realtime PCR Quantification
  Statistical Analysis
 RESULTS
 DISCUSSION
 REFERENCES

저자정보

  • Mi-Rung Park Division of Animal Biotechnology, National Institute of Animal Science, RDA
  • In-Sun Hwang Division of Animal Biotechnology, National Institute of Animal Science, RDA
  • Joo-Hyun Shim Division of Animal Biotechnology, National Institute of Animal Science, RDA
  • Hyo-Jin Moon Division of Animal Biotechnology, National Institute of Animal Science, RDA
  • Dong-Hoon Kim Division of Animal Biotechnology, National Institute of Animal Science, RDA
  • Yeoung-Kyu Ko Division of Animal Biotechnology, National Institute of Animal Science, RDA
  • Hwan-Hoo Seong Division of Animal Biotechnology, National Institute of Animal Science, RDA
  • Gi-Sun Im Division of Animal Biotechnology, National Institute of Animal Science, RDA

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