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The Exocytosis of Fluorescent Nanodiamond and Its Use as a Long-Term Cell Tracker

Authors

  • Chia-Yi Fang,

    1. Institute of Atomic and Molecular Sciences, Academia Sinica Taipei 106, Taiwan
    2. Department of Chemistry, National Taiwan University, Taipei 106, Taiwan
    Current affiliation:
    1. These authors contributed equally to this work.
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  • V. Vaijayanthimala,

    1. Institute of Atomic and Molecular Sciences, Academia Sinica Taipei 106, Taiwan
    2. Molecular Science and Technology Program, Taiwan International Graduate Program, Institute of Atomic and Molecular Sciences, Academia Sinica, Taipei 106, Taiwan
    3. Department of Chemistry, National Tsing Hua University, Hsinchu 300, Taiwan
    Current affiliation:
    1. These authors contributed equally to this work.
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  • Chi-An Cheng,

    1. Institute of Atomic and Molecular Sciences, Academia Sinica Taipei 106, Taiwan
    2. Department of Chemistry, National Taiwan University, Taipei 106, Taiwan
    Current affiliation:
    1. These authors contributed equally to this work.
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  • Shih-Hua Yeh,

    1. Institute of Atomic and Molecular Sciences, Academia Sinica Taipei 106, Taiwan
    2. Department of Chemistry, National Taiwan University, Taipei 106, Taiwan
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  • Ching-Fang Chang,

    1. Genomics Research Center, Academia Sinica Taipei 115, Taiwan
    2. Institute of Microbiology and Immunology, National Yang-Ming University, Taipei 112, Taiwan
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  • Chung-Leung Li,

    1. Genomics Research Center, Academia Sinica Taipei 115, Taiwan
    2. Institute of Cellular and Organismic Biology, Academia Sinica Taipei 115, Taiwan
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  • Huan-Cheng Chang

    Corresponding author
    1. Institute of Atomic and Molecular Sciences, Academia Sinica Taipei 106, Taiwan
    2. Department of Chemistry, National Taiwan University, Taipei 106, Taiwan
    3. Genomics Research Center, Academia Sinica Taipei 115, Taiwan
    • Institute of Atomic and Molecular Sciences, Academia Sinica Taipei 106, Taiwan.
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Abstract

Fluorescent nanodiamond (FND) has excellent biocompatibility and photostability, making it well suited for long-term labeling and tracking of cancer and stem cells. To prove the concept, the exocytosis of FND particles (size ≈100 nm) from three cell lines—HeLa cervical cancer cells, 3T3-L1 pre-adipocytes, and 489-2.1 multipotent stromal cells—is studied in detail. FND labeling is performed by incubating the cells in a serum-free medium containing 80 μg mL−1 FND for 4 h. No significant alteration in growth or proliferation of the FND-labeled cells, including the multipotent stromal cells, is observed for up to 8 days. Flow cytometric analysis, in combination with parallel cell doubling-time measurements, indicates that there is little (≈15% or less) excretion of the endocytosed FND particles after 6 days of labeling for both HeLa and 489-2.1 cells, but exocytosis occurs more readily (up to 30%) for 3T3-L1 preadipocytes. A comparative experiment with FND and the widely used dye, carboxyfluorescein diacetate succinimidyl ester, demonstrates that the nanoparticle platform is a promising alternate probe for long-term cell labeling and tracking applications.

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