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Reprogramming in vivo produces teratomas and iPS cells with totipotency features

Nature volume 502, pages 340345 (17 October 2013) | Download Citation

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Abstract

Reprogramming of adult cells to generate induced pluripotent stem cells (iPS cells) has opened new therapeutic opportunities; however, little is known about the possibility of in vivo reprogramming within tissues. Here we show that transitory induction of the four factors Oct4, Sox2, Klf4 and c-Myc in mice results in teratomas emerging from multiple organs, implying that full reprogramming can occur in vivo. Analyses of the stomach, intestine, pancreas and kidney reveal groups of dedifferentiated cells that express the pluripotency marker NANOG, indicative of in situ reprogramming. By bone marrow transplantation, we demonstrate that haematopoietic cells can also be reprogrammed in vivo. Notably, reprogrammable mice present circulating iPS cells in the blood and, at the transcriptome level, these in vivo generated iPS cells are closer to embryonic stem cells (ES cells) than standard in vitro generated iPS cells. Moreover, in vivo iPS cells efficiently contribute to the trophectoderm lineage, suggesting that they achieve a more plastic or primitive state than ES cells. Finally, intraperitoneal injection of in vivo iPS cells generates embryo-like structures that express embryonic and extraembryonic markers. We conclude that reprogramming in vivo is feasible and confers totipotency features absent in standard iPS or ES cells. These discoveries could be relevant for future applications of reprogramming in regenerative medicine.

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Change history

  • 16 October 2013

    Scale bar values were added to Fig. 5a.

Accessions

Gene Expression Omnibus

Data deposits

The primary RNA-seq data has been deposited in the GEO repository under accession number GSE48364.

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Acknowledgements

We are grateful to M.Torres for advice, and to K. Hochedlinger and R. Jaenisch for reagents. We also thank F. Beier, R. Serrano and N. Soberón for technical support. Work in the laboratory of M.S. is funded by the CNIO and by grants from the Spanish Ministry of Economy (MINECO, SAF), the Regional Government of Madrid (ReCaRe), the European Union (RISK-IR), the European Research Council (ERC Advanced Grant), the Botin Foundation, the Ramon Areces Foundation and the AXA Foundation. Work in the laboratory of M.M. is funded by grants from the MINECO (BFU), the Regional Government of Madrid (Cell-DD) and the ProCNIC Foundation. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Author information

Affiliations

  1. Tumour Suppression Group, Spanish National Cancer Research Centre (CNIO), Madrid E-28029, Spain

    • María Abad
    • , Lluc Mosteiro
    • , Cristina Pantoja
    •  & Manuel Serrano
  2. Histopathology Unit, Spanish National Cancer Research Centre (CNIO), Madrid E-28029, Spain

    • Marta Cañamero
  3. Cardiovascular Development and Repair Department, Spanish National Cardiovascular Research Centre (CNIC), Madrid E-28029, Spain

    • Teresa Rayon
    • , Inmaculada Ors
    •  & Miguel Manzanares
  4. Bioinformatics Unit, Spanish National Cancer Research Centre (CNIO), Madrid E-28029, Spain

    • Osvaldo Graña
  5. Confocal Microscopy Unit, Spanish National Cancer Research Centre (CNIO), Madrid E-28029, Spain

    • Diego Megías
  6. Genomics Unit, Spanish National Cancer Research Centre (CNIO), Madrid E-28029, Spain

    • Orlando Domínguez
  7. Flow Cytometry Unit, Spanish National Cancer Research Centre (CNIO), Madrid E-28029, Spain

    • Dolores Martínez
  8. Transgenic Mice Unit, Spanish National Cancer Research Centre (CNIO), Madrid E-28029, Spain

    • Sagrario Ortega

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Contributions

M.A. performed most of the experiments, contributed to experimental design, data analysis, discussion and writing; L.M. performed a substantial amount of experimental work, contributed to experimental design, data analysis, discussion and writing; C.P. contributed to experimental work, data analysis, discussion and writing; M.C. performed all the histopathological and immunohistochemical analyses; T.R. and I.O. contributed to the trophoblast stem cell and giant cell differentiation assays; O.G. analysed the RNAseq data; D. Megías supervised and helped with the confocal microscopy; O.D. performed RNAseq and determined the lentiviral genomic insertion sites; D. Martínez performed cell sorting and contributed to the bone marrow and peripheral blood analyses; M.M. supervised trophoblast differentiation assays and gave advice; S.O. generated the transgenic mice, constructed chimeras, and perfomed morula and blastocyst assays; M.S. designed and supervised the study, secured funding, analysed the data, and wrote the manuscript. All authors discussed the results and commented on the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Manuel Serrano.

Extended data

Supplementary information

Excel files

  1. 1.

    Supplementary Table 1

    This table shows differentially expressed genes in in vivo iPS cells vs in vitro iPS cells.

  2. 2.

    Supplementary Table 2

    This table shows differentially expressed genes in in vivo iPS cells vs ES cells.

  3. 3.

    Supplementary Table 3

    This table shows differentially expressed genes in ES cells vs in vitro iPS cells.

  4. 4.

    Supplementary Table 4

    This table shows upregulated and downregulated genes in in vivo iPS cells and ES cells (vs in vitro iPS cells).

  5. 5.

    Supplementary Table 5

    This table shows upregulated and downregulated genes in in vivo iPS cells (vs in vitro iPS cells and ES cells).

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DOI

https://doi.org/10.1038/nature12586

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