Adult mice generated from induced pluripotent stem cells

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Abstract

Recent landmark experiments have shown that transient overexpression of a small number of transcription factors can reprogram differentiated cells into induced pluripotent stem (iPS) cells that resemble embryonic stem (ES) cells1,2,3,4,5,6,7. These iPS cells hold great promise for medicine because they have the potential to generate patient-specific cell types for cell replacement therapy and produce in vitro models of disease, without requiring embryonic tissues or oocytes8,9,10. Although current iPS cell lines resemble ES cells, they have not passed the most stringent test of pluripotency by generating full-term or adult mice in tetraploid complementation assays3,11, raising questions as to whether they are sufficiently potent to generate all of the cell types in an organism. Whether this difference between iPS and ES cells reflects intrinsic limitations of direct reprogramming is not known. Here we report fertile adult mice derived entirely from iPS cells that we generated by inducible genetic reprogramming of mouse embryonic fibroblasts. Producing adult mice derived entirely from a reprogrammed fibroblast shows that all features of a differentiated cell can be restored to an embryonic level of pluripotency without exposure to unknown ooplasmic factors. Comparing these fully pluripotent iPS cell lines to less developmentally potent lines may reveal molecular markers of different pluripotent states. Furthermore, mice derived entirely from iPS cells will provide a new resource to assess the functional and genomic stability of cells and tissues derived from iPS cells, which is important to validate their utility in cell replacement therapy and research applications.

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Figure 1: Generation of iPS cells.
Figure 2: Adult mice derived from iMZ cell lines.
Figure 3: Genetic analyses of iPS mice.

References

  1. 1

    Takahashi, K. et al. Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell 131, 861–872 (2007)

  2. 2

    Takahashi, K. & Yamanaka, S. Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell 126, 663–676 (2006)

  3. 3

    Wernig, M. et al. In vitro reprogramming of fibroblasts into a pluripotent ES-cell-like state. Nature 448, 318–324 (2007)

  4. 4

    Park, I. H., Lerou, P. H., Zhao, R., Huo, H. & Daley, G. Q. Generation of human-induced pluripotent stem cells. Nature Protocols 3, 1180–1186 (2008)

  5. 5

    Yu, J. et al. Induced pluripotent stem cell lines derived from human somatic cells. Science 318, 1917–1920 (2007)

  6. 6

    Maherali, N. et al. Directly reprogrammed fibroblasts show global epigenetic remodeling and widespread tissue contribution. Cell Stem Cell 1, 55–70 (2007)

  7. 7

    Zhou, H. et al. Generation of induced pluripotent stem cells using recombinant proteins. Cell Stem Cell 4, 381–384 (2009)

  8. 8

    Ebert, A. D. et al. Induced pluripotent stem cells from a spinal muscular atrophy patient. Nature 457, 277–280 (2009)

  9. 9

    Park, I. H. et al. Disease-specific induced pluripotent stem cells. Cell 134, 877–886 (2008)

  10. 10

    Dimos, J. T. et al. Induced pluripotent stem cells generated from patients with ALS can be differentiated into motor neurons. Science 321, 1218–1221 (2008)

  11. 11

    Hanna, J. et al. Direct reprogramming of terminally differentiated mature B lymphocytes to pluripotency. Cell 133, 250–264 (2008)

  12. 12

    Eggan, K. et al. Mice cloned from olfactory sensory neurons. Nature 428, 44–49 (2004)

  13. 13

    Wakayama, T., Perry, A. C., Zuccotti, M., Johnson, K. R. & Yanagimachi, R. Full-term development of mice from enucleated oocytes injected with cumulus cell nuclei. Nature 394, 369–374 (1998)

  14. 14

    Nagy, A., Rossant, J., Nagy, R., Abramow-Newerly, W. & Roder, J. C. Derivation of completely cell culture-derived mice from early-passage embryonic stem cells. Proc. Natl Acad. Sci. USA 90, 8424–8428 (1993)

  15. 15

    Eggan, K. & Jaenisch, R. in Mammalian and Avian Transgenesis—New Approaches (eds Pease, S. & Lois, C.) Ch. 3 41–68 (Springer, 2006)

  16. 16

    Chin, M. H. et al. Induced pluripotent stem cells and embryonic stem cells are distinguished by gene expression signatures. Cell Stem Cell 5, 111–123 (2009)

  17. 17

    Novak, A., Gup, C., Yang, W., Nagy, A. & Lobe, C. G. Z/EG, a double reporter mouse line that expresses enhanced green fluorescent protein upon cre-mediated excision. Genesis 28, 147–155 (2000)

  18. 18

    Nagai, Y., Sano, H. & Yokoi, M. Transgenic expression of Cre recombinase in mitral/tufted cells of the olfactory bulb. Genesis 43, 12–16 (2005)

  19. 19

    Lois, C., Hong, E. J., Pease, S., Brown, E. J. & Baltimore, D. Germline transmission and tissue-specific expression of transgenes delivered by lentiviral vectors. Science 295, 868–872 (2002)

  20. 20

    Go, W. Y. & Ho, S. N. Optimization and direct comparison of the dimerizer and reverse tet transcriptional control systems. J. Gene Med. 4, 258–270 (2002)

  21. 21

    Yusa, K., Rad, R., Takeda, J. & Bradley, A. Generation of transgene-free induced pluripotent mouse stem cells by the piggyBac transposon. Nature Methods 6, 363–369 (2009)

  22. 22

    Huangfu, D. et al. Induction of pluripotent stem cells by defined factors is greatly improved by small-molecule compounds. Nature Biotechnol. 26, 795–797 (2008)

  23. 23

    Yamanaka, S. Elite and stochastic models for induced pluripotent stem cell generation. Nature 460, 49–52 (2009)

  24. 24

    Mikkelsen, T. S. et al. Dissecting direct reprogramming through integrative genomic analysis. Nature 454, 49–55 (2008)

  25. 25

    Eakin, G. S., Hadjantonakis, A. K., Papaioannou, V. E. & Behringer, R. R. Developmental potential and behavior of tetraploid cells in the mouse embryo. Dev. Biol. 288, 150–159 (2005)

  26. 26

    Eggan, K. & Jaenisch, R. Differentiation of F1 embryonic stem cells into viable male and female mice by tetraploid embryo complementation. Methods Enzymol. 365, 25–39 (2003)

  27. 27

    Mackay, G. E. & West, J. D. Fate of tetraploid cells in 4n↔2n chimeric mouse blastocysts. Mech. Dev. 122, 1266–1281 (2005)

  28. 28

    Wernig, M. et al. A drug-inducible transgenic system for direct reprogramming of multiple somatic cell types. Nature Biotechnol. 26, 916–924 (2008)

  29. 29

    Nagy, A., Gertsenstein, M., Vintersten, K. & Behringer, R. Manipulating the Mouse Embryo: a Laboratory Manual 453–506 (Cold Spring Harbor Laboratory Press, 2003)

  30. 30

    Goebbels, S. et al. Genetic targeting of principal neurons in neocortex and hippocampus of NEX-Cre mice. Genesis 44, 611–621 (2006)

  31. 31

    Wu, S. X. et al. Pyramidal neurons of upper cortical layers generated by NEX-positive progenitor cells in the subventricular zone. Proc. Natl Acad. Sci. USA 102, 17172–17177 (2005)

  32. 32

    Xu, J., Mashimo, T. & Sudhof, T. C. Synaptotagmin-1, -2, and -9: Ca2+ sensors for fast release that specify distinct presynaptic properties in subsets of neurons. Neuron 54, 567–581 (2007)

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Acknowledgements

We wish to thank S. Carlson for help with animal husbandry, K. Spencer for help with imaging, and D. Trajkovic for help with histology. We thank G. Joyce, U. Mueller, L. Stowers, A. Patapoutian and A. Maximov for critical reading of the manuscript. We thank M. Mayford for the gift of rtTAM2.2, A. Maximov for the lentiviral vector backbones, and members of A. Kralli’s laboratory for assistance with qPCR. We thank R. Axel for supporting the generation of the Pcdh21/Cre mouse strain, which was a gift. This work was supported by a Pew Scholars Award (K.K.B.) and grants from the California Institute of Regenerative Medicine, the Whitehall Foundation, the O’Keefe Foundation, and the Shapiro Family Foundation.

Author Contributions M.J.B., J.L.H. and K.L.N. designed and performed experiments, analysed data and edited the manuscript. A.R.R. performed blastocyst injections, Caesarean sections and cross fostering. W.G. assisted in designing and generating lentiviral constructs. G.M. assisted with cell culture. S.K. designed and performed experiments, analysed data and edited the manuscript. K.K.B. conceived of the experimental design, performed experiments, analysed data and wrote the manuscript.

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Correspondence to Kristopher L. Nazor or Kristin K. Baldwin.

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Boland, M., Hazen, J., Nazor, K. et al. Adult mice generated from induced pluripotent stem cells. Nature 461, 91–94 (2009) doi:10.1038/nature08310

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