Article | Published:

Tracking the engraftment and regenerative capabilities of transplanted lung stem cells using fluorescent nanodiamonds

Nature Nanotechnology volume 8, pages 682689 (2013) | Download Citation

Abstract

Lung stem/progenitor cells are potentially useful for regenerative therapy, for example in repairing damaged or lost lung tissue in patients. Several optical imaging methods and probes have been used to track how stem cells incorporate and regenerate themselves in vivo over time. However, these approaches are limited by photobleaching, toxicity and interference from background tissue autofluorescence. Here we show that fluorescent nanodiamonds, in combination with fluorescence-activated cell sorting, fluorescence lifetime imaging microscopy and immunostaining, can identify transplanted CD45CD54+CD157+ lung stem/progenitor cells in vivo, and track their engraftment and regenerative capabilities with single-cell resolution. Fluorescent nanodiamond labelling did not eliminate the cells’ properties of self-renewal and differentiation into type I and type II pneumocytes. Time-gated fluorescence imaging of tissue sections of naphthalene-injured mice indicates that the fluorescent nanodiamond-labelled lung stem/progenitor cells preferentially reside at terminal bronchioles of the lungs for 7 days after intravenous transplantation.

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Acknowledgements

This work was supported by Academia Sinica and the National Science Council, Taiwan (grants nos. 100-2119M-001-028 and 100-2321-B-001-036). The authors thank the Division of Medical Biology, Genomics Research Center, for providing facilities and support for cell imaging and flow cytometric analysis.

Author information

Author notes

    • Tsai-Jung Wu
    •  & Yan-Kai Tzeng

    These authors contributed equally to this work

Affiliations

  1. Institute of Biochemistry and Molecular Biology, Program in Molecular Medicine, School of Life Sciences, National Yang-Ming University, Taipei 112, Taiwan

    • Tsai-Jung Wu
    • , Chin-Hsiang Chien
    •  & John Yu
  2. Genomics Research Center, Academia Sinica, Taipei 115, Taiwan

    • Tsai-Jung Wu
    • , Wei-Wei Chang
    • , Huan-Cheng Chang
    •  & John Yu
  3. Institute of Cellular and Organismic Biology, Academia Sinica, Taipei 115, Taiwan

    • Tsai-Jung Wu
    • , Wei-Wei Chang
    •  & John Yu
  4. Institute of Atomic and Molecular Sciences, Academia Sinica, Taipei 106, Taiwan

    • Yan-Kai Tzeng
    • , Chi-An Cheng
    • , Yung Kuo
    •  & Huan-Cheng Chang
  5. Department of Chemistry, National Taiwan University, Taipei 106, Taiwan

    • Yan-Kai Tzeng
    • , Yung Kuo
    •  & Huan-Cheng Chang
  6. Center of Stem Cells and Translational Cancer Research, Chang Gung Memorial Hospital, Linkou, Taoyuan County 333, Taiwan

    • John Yu

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Contributions

T.J.W. and Y.K.T. conceived the study, conducted the experiments, interpreted the results, and wrote the manuscript. T.J.W. performed the tissue-culture work, cell sorting and tracking experiments, and data analysis. Y.K.T. and Y.K. assembled the FLIM systems and produced the image acquisition scripts. W.W.C. helped with the isolation of LSCs and carried out data analysis. C.A.C. prepared FNDs and performed the cell tracking experiments. C.H.C., H.C.C. and J.Y. supervised and planned the project, analysed the data, and wrote the manuscript. All authors designed the experiments and edited the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Huan-Cheng Chang or John Yu.

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DOI

https://doi.org/10.1038/nnano.2013.147

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