Tracking the engraftment and regenerative capabilities of transplanted lung stem cells using fluorescent nanodiamonds.
Tsai-Jung Wu,Yan-Kai Tzeng,Yan-Kai Tzeng,Wei-Wei Chang,Chi-An Cheng,Yung Kuo,Yung Kuo,Chin-Hsiang Chien,Huan-Cheng Chang,Huan-Cheng Chang,John Yu +10 more
TLDR
It is shown that fluorescent nanodiamonds, in combination with fluorescence-activated cell sorting, fluorescence lifetime imaging microscopy and immunostaining, can identify transplanted lung stem/progenitor cells in vivo, and track their engraftment and regenerative capabilities with single-cell resolution.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 CD45(-)CD54(+)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.read more
Citations
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Mass production and dynamic imaging of fluorescent nanodiamonds
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TL;DR: It is shown that bright fluorescent nanodiamonds can be produced in large quantities by irradiating synthetic diamond nanocrystallites with helium ions, and the fluorescence is sufficiently bright and stable to allow three-dimensional tracking of a single particle within the cell by means of either one- or two-photon-excited fluorescence microscopy.
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