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Related Experiment Videos

Cloning sheep from cultured embryonic cells.

D N Wells1, P M Misica, A M Day

  • 1Reproductive Technologies Group, AgResearch Ruakura, Hamilton, New Zealand. wellsd@agresearch.cri.nz

Reproduction, Fertility, and Development
|December 28, 1999
PubMed
Summary

Cloned lambs were successfully produced from established embryonic cell lines using nuclear transfer. While improvements were noted, the process remains inefficient, requiring further research for healthier fetuses.

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Area of Science:

  • Reproductive biology
  • Animal cloning
  • Developmental biology

Background:

  • Totipotent embryonic cell lines are crucial for efficient transgenic animal production.
  • Nuclear transfer (NT) is a key technology for animal cloning.

Purpose of the Study:

  • To generate cloned lambs from established embryonic cell lines using NT.
  • To evaluate factors affecting NT efficiency, including oocyte source and co-culture methods.

Main Methods:

  • Nuclear transfer using cytoplasts from in vivo and in vitro oocytes.
  • Co-culture of cloned embryos with oviductal epithelial cells or synthetic oviductal fluid.
  • Transfer of cloned embryos with and without trophoblastic vesicles.
  • Monitoring embryo development, pregnancy, and term survival.

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Main Results:

  • Cloned male and female lambs were produced from two embryonic cell lines.
  • In vivo oocytes yielded higher blastocyst development (24%) and term survival (7%) compared to in vitro oocytes (17% and 2%).
  • Co-culture methods showed no significant advantage over synthetic oviductal fluid.
  • Trophoblastic vesicle co-transfer improved Day 35 survival but not term survival.
  • Significant fetal loss occurred in the last trimester, with an overall term survival rate of 11%.

Conclusions:

  • Established embryonic cell lines can produce cloned lambs via NT.
  • In vivo oocytes are superior to in vitro oocytes for NT in this model.
  • Significant inefficiencies persist in the NT process, particularly during embryo attachment and late-term gestation.
  • Further research is essential to enhance the efficiency and health outcomes of cloned animals.