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Cloning Mice and ES Cells by Nuclear Transfer from Somatic Stem Cells and Fully Differentiated Cells

Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 770)

Abstract

Cloning animals by nuclear transfer (NT) has been successful in several mammalian species. In addition to cloning live animals (reproductive cloning), this technique has also been used in several species to establish cloned embryonic stem (ntES) cell lines from somatic cells. It is the latter application of this technique that has been heralded as being the potential means to produce isogenic embryonic stem cells from patients for cell therapy (therapeutic cloning). These two types of cloning differ only in the steps after cloned embryos are produced: for reproductive cloning the cloned embryos are transferred to surrogate mothers to allow them to develop to full term and for therapeutic cloning the cloned embryos are used to derive ntES cells. In this chapter, a detailed NT protocol in mouse by using somatic stem cells (neuron and skin stem cells) and fully differentiated somatic cells (cumulus cells and fibroblast cells) as nuclear donors is described.

Key words

Nuclear transfer animal cloning epigenetic reprogramming ES cell ntES cell somatic stem cell oocyte embryo 

Notes

Acknowledgments

The work is supported by Hematech, Inc. The author thanks Emma Wang from Harvard College for proofreading the manuscript.

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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  1. 1.Department of Epigenetics and Embryo DevelopmentHematech, Inc.Sioux FallsUSA

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