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A Novel Method of Somatic Cell Nuclear Transfer with Minimum Equipment

  • S. M. HosseiniEmail author
  • F. Moulavi
  • M. H. Nasr-EsfahaniEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1330)

Abstract

Somatic cell nuclear transfer (SCNT) is an exceptional experimental biology technique with an arguably great contribution to our current understanding of developmental plasticity. Many students and young researchers are interested in taking advantage of SCNT virtues in their experiments but the cost of micromanipulation microscopes, intensive training programs, and also the sophisticated process of SCNT may dissuade them from entering this amazing field of science. Here, we describe the details of a streamlined manual method of SCNT that can be performed using very basic equipment found in every embryology laboratory: the Pasteur pipette and stereomicroscope. The overall method introduced is very simple and a person with no previous experience in cloning can learn and adopt the basic routines of this technique independently.

Key words

Somatic cell nuclear transfer Manual method Pasteur pipette 

Notes

Acknowledgment

This study was supported by a Royan Institute grant for the Advancement of Reproductive Biotechnology. The Authors would like to thank the staff of the Embryology Department of Royan Institute for kind help in this project.

Supplementary material

Video 1

Blind manual oocyte enucleation (BME) (MP4 2167 kb)

Video 2

Oriented manual oocyte enucleation (OME) (MP4 3396 kb)

References

  1. 1.
    Spemann H (1938) Embryonic development and induction. Hafner Publishing Co, New YorkGoogle Scholar
  2. 2.
    Nasr-Esfahani MH, Hosseini SM, Hajian M et al (2011) Development of an optimized zona-free method of somatic cell nuclear transfer in the goat. Cell Reprogram 13:157–170CrossRefPubMedGoogle Scholar
  3. 3.
    Hosseini SM, Hajian M, Moulavi F et al (2013) Cloned sheep blastocysts derived from oocytes enucleated manually using a pulled pasteur pipette. Cell Reprogram 15:15–23PubMedGoogle Scholar
  4. 4.
    Khoub Bakht Z, Hajian M, Hosseini M et al (2008) Effect of demecolicne treatment on the developmental competency of in vitro matured bovine oocytes. J Iran Anat Sci 6:447–457Google Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  1. 1.Department of Reproductive Biotechnology at Reproductive Biomedicine Research CenterRoyan Institute for Biotechnology, ACECRIsfahanIran

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