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Original Article

Efficient method for generating homozygous embryonic stem cells in mice

초록

영어

Parthenogenesis is maternally uniparental reproduction through the embryonic development of oocytes without fertilization. Artificial activation of mature oocytes could generate homozygous haploid embryos with the extrusion of the second polar body. However, the haploid embryos showed low embryo development in preimplantation embryos. In this study, we investigated whether the electronic fusion of the haploid embryos could enhance embryo development and ESC establishment in mice. Haploid embryos showed the developmental delay from 4-cell to the blastocyst stage. The haploid blastomeres of the 2-cell stage were fused electronically, resulting in that the fused embryos showed a significantly higher rate of blastocysts compared to non-fused haploid embryos (55% vs. 37%). Further, the embryonic stem cells (ESCs) derived from the fused embryos were confirmed to be diploid. The rate of ESC establishment in fused embryos was significantly higher compared to non-fused ones. Based on the results, we concluded that the electronic fusion of haploid embryos could be efficient to generate homozygous ESCs.

목차

ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
Animals
Metaphase II (MII) oocytes collection
Activation of oocytes and embryo culture
Electronic fusion
Establishment and culture of mESCs
Cell cycle analysis
Karyotyping
Statistical analyses
RESULTS
DISCUSSION
CONCLUSION
REFERENCES

저자정보

  • Bitnara Kim Department of Biomedical Science, College of Life Science, CHA University, Seongnam 13488, Korea
  • Seongjun So Department of Biomedical Science, College of Life Science, CHA University, Seongnam 13488, Korea
  • Jiwan Choi Center for Embryo and Stem Cell Research, CHA Advanced Research Institute, CHA University, Seongnam 13488, Korea
  • Eunju Kang Department of Biomedical Science, College of Life Science, CHA University, Center for Embryo and Stem Cell Research, CHA Advanced Research Institute, CHA University, Seongnam 13488, Korea
  • Yeonmi Lee Department of Biomedical Science, College of Life Science, CHA University, Center for Embryo and Stem Cell Research, CHA Advanced Research Institute, CHA University, Seongnam 13488, Korea

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