Abstract

Although cellular therapies represent a promising strategy for a number of conditions, current approaches face major translational hurdles, including limited cell sources and the need for cumbersome pre-processing steps (for example, isolation, induced pluripotency)1,2,3,4,5,6. In vivo cell reprogramming has the potential to enable more-effective cell-based therapies by using readily available cell sources (for example, fibroblasts) and circumventing the need for ex vivo pre-processing7,8. Existing reprogramming methodologies, however, are fraught with caveats, including a heavy reliance on viral transfection9,10. Moreover, capsid size constraints and/or the stochastic nature of status quo approaches (viral and non-viral) pose additional limitations, thus highlighting the need for safer and more deterministic in vivo reprogramming methods11,12. Here, we report a novel yet simple-to-implement non-viral approach to topically reprogram tissues through a nanochannelled device validated with well-established and newly developed reprogramming models of induced neurons and endothelium, respectively. We demonstrate the simplicity and utility of this approach by rescuing necrotizing tissues and whole limbs using two murine models of injury-induced ischaemia.

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Acknowledgements

Funding for C.K.S. was partly provided by NIGMS/NINR (R01GM07718507, R01GM10801402, R01NR01567601, R01NR01389804 and R01NS42617) and a philanthropic gift from Leslie and Abigail Wexner. Funding for L.J.L. was partly provided by NIBIB (R21EB017539), NSF (NSEC EEC-0914790) and the National Center for the Advancing Translational Sciences (UL1TR001070). Funding for D.G.-P., S.K. and C.R. was partly provided by NINDS (R21NS099869). Additional funding for D.G.-P. and S.K. was provided in part by the NIDDK Diabetic Complications Consortium (DiaComp, www.diacomp.org), grant DK076169 (U24DK076169). J.J.O., V.B.S., S.K., C.R. and S.R. acknowledge financial support from NIH (R01HL132355, R21EB017539, R01NS099869 and R01DK076566) and NSF (1325114), respectively. The authors thank T. Wilgus and B. Wulff (Department of Pathology, The Ohio State University) for providing the Col1A1 mice used in this study. Fsp1-Cre mice were a gift from A. Deb (University of California, Los Angeles). Etv2 and Fli1 plasmids were donated by A. Ferdous (Department of Internal Medicine, UT Southwestern). Foxc2 plasmid was donated by T. Kume (Department of Medicine-Cardiology and Pharmacology, Northwestern University-FCVRI, Chicago). X. Wang (The Ohio State University) provided support with plasmid design and preparation. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Center for Advancing Translational Sciences, National Science Foundation or the National Institutes of Health. This work was sponsored by and represents activity of The Ohio State University Center for Regenerative Medicine and Cell Based Therapies (regenerativemedicine.osu.edu) and Nanoscale Engineering Center for Affordable Nanoengineering of Polymeric Biomedical Devices.

Author information

Author notes

    • Daniel Gallego-Perez
    • , Durba Pal
    •  & Subhadip Ghatak

    These authors contributed equally to this work.

Affiliations

  1. Department of Surgery, The Ohio State University, Columbus, Ohio 43210, USA

    • Daniel Gallego-Perez
    • , Durba Pal
    • , Subhadip Ghatak
    • , Natalia Higuita-Castro
    • , Surya Gnyawali
    • , Mithun Sinha
    • , Kanhaiya Singh
    • , Erin Steen
    • , Alec Sunyecz
    • , Richard Stewart
    • , Jordan Moore
    • , Sashwati Roy
    • , Savita Khanna
    • , Cameron Rink
    •  & Chandan K. Sen
  2. Department of Biomedical Engineering, The Ohio State University, Columbus, Ohio 43210, USA

    • Daniel Gallego-Perez
    •  & Lingqian Chang
  3. Center for Affordable Nanoengineering of Polymeric Biomedical Devices, The Ohio State University, Columbus, Ohio 43210, USA

    • Daniel Gallego-Perez
    • , Veysi Malkoc
    • , Lingqian Chang
    • , Wei-Ching Liao
    • , Junfeng Shi
    •  & L. James Lee
  4. Center for Regenerative Medicine and Cell-Based Therapies, The Ohio State University, Columbus, Ohio 43210, USA

    • Daniel Gallego-Perez
    • , Durba Pal
    • , Subhadip Ghatak
    • , Natalia Higuita-Castro
    • , Surya Gnyawali
    • , Mithun Sinha
    • , Kanhaiya Singh
    • , Alec Sunyecz
    • , Richard Stewart
    • , Jordan Moore
    • , Sashwati Roy
    • , Savita Khanna
    • , Cameron Rink
    • , Jose J. Otero
    • , L. James Lee
    •  & Chandan K. Sen
  5. Department of Chemical and Biomolecular Engineering, The Ohio State University, Columbus, Ohio 43210, USA

    • Veysi Malkoc
    • , Alec Sunyecz
    • , Michael Homsy
    •  & L. James Lee
  6. Department of Mechanical and Aerospace Engineering, The Ohio State University, Columbus, Ohio 43210, USA

    • Junfeng Shi
    • , Thomas Ziebro
    • , Robert G. Northcutt
    •  & Vishnu Baba Sundaresan
  7. Department of Electrical and Computer Engineering, The Ohio State University, Columbus, Ohio 43210, USA

    • Paul Bertani
    •  & Wu Lu
  8. Department of Pathology, The Ohio State University, Columbus, Ohio 43210, USA

    • Jose J. Otero
  9. Department of Neuroscience, The Ohio State University, Columbus, Ohio 43210, USA

    • Jose J. Otero

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Contributions

TNT platform design, implementation and optimization (for different applications) was performed by D.G.-P., L.J.L., D.P., S.G. and C.K.S. TNT chip fabrication, transductions, cell/tissue imaging, transgenic mouse experiments and histology were performed by D.G.-P., D.P., S.G., N.H.-C., V.M., S.G., L.C., M.S., E.S., A.S., J.M., P.B., W.L., J.J.O., L.J.L. and C.K.S. Electrophysiological activity measurements were conducted by T.Z., R.G.N., V.B.S. and J.J.O., with support from S.G. and N.H.-C. Global gene expression analyses were conducted by S.R., S.K., K.S. and C.K.S. TNT chip simulations were conducted by W.-C.L., J.S., L.C., D.G.-P. and L.J.L. S.R. oversaw and participated in the LCM work. Stroke recovery experiments were conducted by D.G.-P., V.M., A.S., R.S., M.H., S.K., C.R. and C.K.S. The manuscript was written by D.G.-P., L.J.L., D.P., S.G. and C.K.S. C.K.S. and L.J.L. jointly supervised this work.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to L. James Lee or Chandan K. Sen.

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DOI

https://doi.org/10.1038/nnano.2017.134

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