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논문검색

In Vitro Development of Interspecies Nuclear Transfer Embryos: Effects of Culture Systems

초록

영어

Porcine fibroblasts were transferred into enucleated bovine oocytes for the interspecies nuclear transfer (NT). After NT, the embryos were cultured in three different culture systems. The media used for the experiment were CR1aa and NCSU23. The culture systems used for the experiment were: 1. Culture in CR1aa for 7 days (CR). 2. Culture in CR1aa for 2 days and subsequently in NCSU23 for 5 days (CR-NC). 3. Culture in NCSU23 for 7 days (NC). Bovine (intraspecies) NT group was used as a control. The oocytes in bovine NT group were treated the same as interspecies NT embryos except using bovine fibroblasts as nuclear donors. Regardless of their nuclear origin (interspecies vs bovine), the embryos in CR (68.4% vs 77.2%) and CR-NC (67.8% vs 70.5%) showed better developmental competence to the 2-cell stage (p<0.05) than those in NC (41.0% vs 10.0%). Bovine NT embryos in CR-NC did not develop over the 4-cell stage after the medium replacement, while interspecies NT embryos in CR-NC continued to develop and could reach over the 8-cell stage (12.2%). Blastocysts were only found in bovine NT group (17.4%), but no blastocyst was found in interspecies NT group. This study suggests that the development of interspecies NT embryos mostly depends on their recipient cytoplasm during the culture in vitro.

목차

INTRODUCTION
 MATERIALS AND METHODS
  Reagents
  In Vitro Maturation of Bovine Oocytes
  Nuclear Donor Cell Preparation
  Enucleation
  Preparation of Donor Cells
  Nuclear Transfer
  Cell Fusion
  Activation
  Culture of Inter-Species NT Embryos
  Statistic Analysis
 RESULTS
 DISCUSSION
 REFERENCES

저자정보

  • Roh Sangho Section of Oral and Craniofacial Biotechnology Research, Dental Research Institute and College of Dentistry, Seoul National University

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