Institute of Biochemistry and Molecular Biology, Program in Molecular Medicine, School of Life Sciences, National Yang-Ming University, Taipei 112, Taiwan.
Nat Nanotechnol. 2013 Sep;8(9):682-9. doi: 10.1038/nnano.2013.147. Epub 2013 Aug 4.
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.
肺干/祖细胞在再生治疗中具有潜在的应用价值，例如在修复患者受损或丧失的肺组织方面。已经有几种光学成像方法和探针被用于跟踪干细胞在体内随时间推移的整合和自我再生情况。然而，这些方法受到光漂白、毒性和背景组织自发荧光干扰的限制。在这里，我们展示了荧光纳米金刚石与荧光激活细胞分选、荧光寿命成像显微镜和免疫染色相结合，可以在体内识别移植的 CD45(-)CD54(+)CD157(+)肺干/祖细胞，并以单细胞分辨率跟踪它们的植入和再生能力。荧光纳米金刚石标记并没有消除细胞的自我更新和分化为 I 型和 II 型肺泡细胞的特性。对萘损伤小鼠组织切片的时间门控荧光成像表明，荧光纳米金刚石标记的肺干/祖细胞在静脉移植后 7 天内优先存在于终末细支气管中。
