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논문검색

Developmental Characteristics of SCNT Pig Embryos Knocked-out of Alpha-1,3-Galactosyltransferase Gene

초록

영어

This study was performed to comprehend the developmental characteristics of cloned embryos knocked out (KO) of α-1,3-galactosyltransferase (GalT) gene. Immature oocytes were collected and cultured for 40 hrs (1-step) or 20hrs (with hormone) + 20hrs (without hormone) (2-step). The embryos transferred with miniature pig ear fibroblast cell were used as control. The reconstructed embryos were cultured in PZM-3 with 5% CO2 in air at 38.5℃ for 6 days. To determine the quality of the blstocysts, TUNEL and quantitative realtime RT-PCR were performed. The embryos were transferred to a surrogate (Landrace) at an earlier stage of the estrus cycle. The maturation rate was significantly higher in 2-step method than that of 1-step (p<0.05). The blastocyst development of GalT KO embryos was significantly lower than that of normal cloned embryos (p<0.05). The total and apoptotic cell number of GalT KO blastocysts was not different statistically from control. The relative abundance of Bax-α/Bcl-xl ratio was significantly higher in both cloned blastocysts than that of in vivo blastocysts (p<0.05). Taken together, it can be postulated that the lower developmental potential and higher expression of apoptosis related genes in GalT KO SCNT embryos might be a cause of a low efficiency of GalT KO cloned miniature pig production.

목차

ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
  Chemicals
  In Vitro Maturation of Oocytes
  Nuclear Transfer Procedure
  Collection of In Vivo Blastocysts
  TUNEL Assay for Apoptotic Cell Death
  Apoptosis Related Gene Expression
  Statistical Analysis
 RESULTS
  In Vitro Maturation of Porcine Oocytes
  In Vitro Development of SCNT Embryos
  Apoptosis Rate in mNT and GalT KO NT Blastocysts
  Analysis of Apoptosis Gene Expression in Blastocysts
 DISCUSSION
 REFERENCES

저자정보

  • Joo-Hyun Shim1 National Institute of Animal Science, RDA
  • Mi-Rung Park National Institute of Animal Science, RDA
  • Byoung-Chul Yang National Institute of Animal Science, RDA
  • Yeoung-Gyu Ko National Institute of Animal Science, RDA
  • Keon Bong Oh National Institute of Animal Science, RDA
  • Jeong-Woong Lee Center for Regenerative Medicine, KRIBB
  • Jae-Seok Woo National Institute of Animal Science, RDA
  • Eung-Woo Park National Institute of Animal Science, RDA
  • Soo-Bong Park National Institute of Animal Science, RDA
  • Seongsoo Hwang National Institute of Animal Science, RDA

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