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Comparison of Microtubule Distributions between Somatic Cell Nuclear Transfer and Parthenogenetic Porcine Embryos

초록

영어

The aim of this study was to examine the microtubule distributions of somatic cell nuclear transfer (SCNT) and parthenogenetic porcine embryos. Porcine SCNT embryos were produced by fusion of serum-starved fetal fibroblast cells with enucleated oocytes. Reconstituted and mature oocytes were activated by electric pulses combined with 6-dimethlyaminopurine treatment. SCNT and parthenogenetic embryos were cultured in vitro for 6 days. Microtubule assembly of embryos was examined by confocal microscopy 1 hr and 20 hr after fusion or activation, respectively. The proportions of embryos developed to the blastocyst stage were 25.7% and 30.4% in SCNT and parthenogenetic embryos, respectively. The frequency of embryos showing β-tubulins was 81.8% in parthenogenetic embryos, whereas 31.3% in SCNT embryos 1 hr after activation or fusion. The frequency of the embryos underwent normal mitotic phase was low in SCNT embryos (40.6%) compared to that of parthenogenetic ones (59.7%) 20 hr after fusion or activation (p<0.05). The rate of SCNT embryos with an abnormal mitosis pattern is about twice compared to that of parthenogenetic ones. The spindle assembly and its distribution of SCNT embryos in the first mitotic phase were not different from those of parthenogenetic ones. The result shows that although microtubule distribution of porcine SCNT embryos shortly after fusion is different from parthenogenetic embryos, and the frequency of abnormal mitosis 20 hr after fusion or activation is slightly increased in SCNT embryos, microtubule distributions at the first mitotic phase are similar in both SCNT and parthenogenetic embryos.

목차

ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
  In Vitro Maturation of Oocytes
  Preparation of Porcine Fetal Fibroblasts
  Micromanipulation
  Electrofusion And Activation
  Parthenogenesis and In Vitro Culture
  Immunofluorescent Staining and Confocal Microscopy
  Statistical Analysis
 RESULTS
  In Vitro Development of  Embryos
  Microtubule Distnbution Shortly after Fusion or Activation
  Microtubule Distribution in the First Mitotic Phase
 DISCUSSION
 REFERENCES

저자정보

  • Joo-Hee Park School of Veterinary Medicine
  • Dae-Jin Kwon School of Veterinary Medicine
  • Beom-Ki Lee School of Veterinary Medicine
  • In-Sun Hwang College of Animal Life Science, Kangwon National University
  • Choon-Keun Park College of Animal Life Science, Kangwon National University
  • Boo-Keun Yang College of Animal Life Science, Kangwon National University
  • Hee-Tae Cheong School of Veterinary Medicine

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