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Pluripotent stem cells induced from adult neural stem cells by reprogramming with two factors

Abstract

Reprogramming of somatic cells is a valuable tool to understand the mechanisms of regaining pluripotency and further opens up the possibility of generating patient-specific pluripotent stem cells. Reprogramming of mouse and human somatic cells into pluripotent stem cells, designated as induced pluripotent stem (iPS) cells, has been possible with the expression of the transcription factor quartet Oct4 (also known as Pou5f1), Sox2, c-Myc and Klf4 (refs 1–11). Considering that ectopic expression of c-Myc causes tumorigenicity in offspring2 and that retroviruses themselves can cause insertional mutagenesis, the generation of iPS cells with a minimal number of factors may hasten the clinical application of this approach. Here we show that adult mouse neural stem cells express higher endogenous levels of Sox2 and c-Myc than embryonic stem cells, and that exogenous Oct4 together with either Klf4 or c-Myc is sufficient to generate iPS cells from neural stem cells. These two-factor iPS cells are similar to embryonic stem cells at the molecular level, contribute to development of the germ line, and form chimaeras. We propose that, in inducing pluripotency, the number of reprogramming factors can be reduced when using somatic cells that endogenously express appropriate levels of complementing factors.

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Figure 1: Generation of two-factor Oct4/Klf4 (OK) iPS cells from adult NSCs of OG2/ROSA26 transgenic mice.
Figure 2: Gene expression profile of iPS cells.
Figure 3: Two-factor Oct4/Klf4 (OK) iPS cells (clone F-4) are pluripotent and differentiate in vitro and in vivo.
Figure 4: In vivo developmental potential of two-factor Oct4/Klf4 (OK) iPS cells (clone F-4).

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Gene Expression Omnibus

Data deposits

The microarray data sets are available from the GEO (Gene Expression Omnibus) website under accession number GSE10806.

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Acknowledgements

We thank J. Müller-Keuker for critically reviewing the manuscript, M. Stehling for fluorescence-activated cell sorting analysis, C. Ortmeier for real-time PCR analysis, B. Schäfer for histology, C. Becker, B. Kratz and B. Denicke for probe processing and array hybridization, and T. Kitamura for the pMX retroviral vector. J.B.K. was supported by the Interdepartmental Graduate-Programme for Experimental Life Sciences (iGEL) at the University of Münster. The microarray analyses were funded in part by a grant from the Deutsche Forschungsgemeinschaft DFG SPP1109.

Author Contributions J.B.K. (project design, generation and characterization of iPS cells and NSCs, and preparation of manuscript), H.Z. (project design, generation of iPS cells, and preparation of manuscript), G.W., L.G., V.S., K.K. and D.W.H. (characterization of iPS cells), M.J.A.-B., D.R., M.Z. (microarray and bioinformatics) and H.R.S. (project design, and preparation of manuscript).

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Correspondence to Hans R. Schöler.

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Kim, J., Zaehres, H., Wu, G. et al. Pluripotent stem cells induced from adult neural stem cells by reprogramming with two factors. Nature 454, 646–650 (2008). https://doi.org/10.1038/nature07061

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