A protocol for adult somatic cell nuclear transfer in medaka fish (Oryzias latipes) with a high rate of viable clone formation. Other Scholarly Work

Bubenshchikova, Ekaterina, Kaftanovskaya, Elena, Adachi, Tomoko et al. (2013). A protocol for adult somatic cell nuclear transfer in medaka fish (Oryzias latipes) with a high rate of viable clone formation. . 15(6), 520-530. 10.1089/cell.2013.0004

cited authors

  • Bubenshchikova, Ekaterina; Kaftanovskaya, Elena; Adachi, Tomoko; Hashimoto, Hisashi; Kinoshita, Masato; Wakamatsu, Yuko

fiu authors

abstract

  • Previously, we successfully generated fully grown, cloned medaka (the Japanese rice fish, Oryzias latipes) using donor nuclei from primary culture cells of adult caudal fin tissue and nonenucleated recipient eggs that were heat shock-treated to induce diploidization of the nuclei. However, the mechanism of clone formation using this method is unknown, and the rate of adult clone formation is not high enough for studies in basic and applied sciences. To gain insight into the mechanism and increase the success rate of this method of clone formation, we tested two distinct nuclear transfer protocols. In one protocol, the timing of transfer of donor nuclei was changed, and in the other, the size of the donor cells was changed; each protocol was based on our original methodology. Ultimately, we obtained an unexpectedly high rate of adult clone formation using the protocol that differed with respect to the timing of donor nuclei transfer. Specifically, 17% of the transplants that developed to the blastula stage ultimately developed into adult clones. The success rate with this method was 13 times higher than that obtained using the original method. Analyses focusing on the reasons for this high success rate of clone formation will help to elucidate the mechanism of clone formation that occurs with this method.

publication date

  • December 1, 2013

keywords

  • Animals
  • Base Sequence
  • Cells, Cultured
  • Clone Cells
  • DNA Primers
  • Female
  • Fluorescence
  • Genetic Markers
  • Male
  • Nuclear Transfer Techniques
  • Oryzias
  • Polymerase Chain Reaction

Digital Object Identifier (DOI)

Medium

  • Print-Electronic

start page

  • 520

end page

  • 530

volume

  • 15

issue

  • 6