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Trends in Stem Cell Biology and Technology pp 101–121Cite as

Observing and Manipulating Pluripotency in Normal and Cloned Mouse Embryos

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Abstract

The mouse ooplasm is the ideal platform to study and compare induced and natural pluripotency because it can support both, after somatic cell nuclear transfer (cloning) and after fertilization, respectively. The amount of pluripotency induced after cloning is variable but always limited compared to fertilization. It can be visualized conveniently if the nucleus donor cells carry a green fluorescent protein (GFP) reporter under control of the pluripotency-associated gene Oct4 promoter. Thus we produced cloned and fertilized mouse embryos transgenic for Oct4-GFP (GOF18-∆PE-EGFP). We also developed and validated a live cell imaging method, whereby we resolve and selectively pick cloned embryos that hold distinct amounts of induced pluripotency as predicted by GFP intensity and measured by embryonic stem cell derivation. Currently we are developing a microinjection method to change the level of Oct4 without modifying the genome of the embryo. Here we discuss our findings in relation to the epigenetic reprogramming of the nucleus transplant and to cell fate decisions in the cloned or fertilized mouse embryo.

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Acknowledgments

We are grateful to Dr. Konstantinos Anastassiadis (Technical University Dresden) for sharing unpublished data on GOF18-ΔPE-EGFP; to Dr. Yong-Mahn Han (KAIST, Korea) for contributing to the article of Cavaleri et al., 2008 (64) which provides the groundwork for this chapter; and to Prof. Ivan Dikic (Goethe University, Frankfurt am Main) for supporting the quantitative analysis of Oct4 protein in preimplantation mouse embryos and the synthesis of recombinant Oct4 for microinjection.

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  1. Max Planck Institute for Molecular Biomedicine, Röntgenstrasse 20, 48149, Münster, Germany

    Sebastian T. Balbach, F. M. Cavaleri, Luca Gentile, Marcos J. Araúzo-Bravo, Hans R. Schöler, N. Crosetto & Michele Boiani

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  1. Sebastian T. Balbach
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  2. F. M. Cavaleri
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  3. Luca Gentile
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  5. Hans R. Schöler
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  7. Michele Boiani
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Correspondence to Michele Boiani .

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    Balbach, S.T. et al. (2009). Observing and Manipulating Pluripotency in Normal and Cloned Mouse Embryos. In: Baharvand, H. (eds) Trends in Stem Cell Biology and Technology. Humana Press. https://doi.org/10.1007/978-1-60327-905-5_7

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