Bioorthogonal Copper Free Click Chemistry for Labeling and Tracking of Chondrocytes In Vivo

  • Bioconjug Chem. 2016 Apr 20;27(4):927-36. doi: 10.1021/acs.bioconjchem.6b00010.
Hwa In Yoon  1  2 ,  Ji Young Yhee  3 ,  Jin Hee Na  4 ,  Sangmin Lee  4 ,  Hyukjin Lee  3 ,  Sun-Woong Kang  5  6 ,  Hyeyoun Chang  1  7 ,  Ju Hee Ryu  1 ,  Seulki Lee  4 ,  Ick Chan Kwon  1 ,  Yong Woo Cho  2 ,  Kwangmeyung Kim  1  7
Affiliations
  • 1. Center for Theragnosis, Biomedical Research Institute, Korea Institute of Science and Technology , Hwarangno 14-gil 6, Seongbuk-gu, Seoul 136-791, Republic of Korea.
  • 2. Departments of Chemical Engineering and Bionanotechnology, Hanyang University , Ansan, Gyeonggi-do 426-791, Republic of Korea.
  • 3. College of Pharmacy, Graduate School of Pharmaceutical Sciences, Ewha Womans University , Seoul 120-750, Republic of Korea.
  • 4. The Russell H. Morgan Department of Radiology and Radiological Sciences, Johns Hopkins University School of Medicine , Baltimore, Maryland United States.
  • 5. Next-generation Pharmaceutical Research Center, Korea Institute of Toxicology , Daejeon 305-343, Republic of Korea.
  • 6. Human and Environmental Toxicology Program, University of Science and Technology (UST) , Daejeon 305-350, Republic of Korea.
  • 7. Department of Biomedical Engineering, University of Science and Technology (UST) , Seoul 136-791, Republic of Korea.
PMID: 26930274 DOI: 10.1021/acs.bioconjchem.6b00010
Abstract

Establishment of an appropriate cell labeling and tracking method is essential for the development of cell-based therapeutic strategies. Here, we are introducing a new method for cell labeling and tracking by combining metabolic gylcoengineering and bioorthogonal copper-free Click chemistry. First, chondrocytes were treated with tetraacetylated N-azidoacetyl-D-mannosamine (Ac4ManNAz) to generate unnatural azide groups (-N3) on the surface of the cells. Subsequently, the unnatural azide groups on the cell surface were specifically conjugated with near-infrared fluorescent (NIRF) dye-tagged dibenzyl cyclooctyne (DBCO-650) through bioorthogonal copper-free Click chemistry. Importantly, DBCO-650-labeled chondrocytes presented strong NIRF signals with relatively low cytotoxicity and the amounts of azide groups and DBCO-650 could be easily controlled by feeding different amounts of Ac4ManNAz and DBCO-650 to the Cell Culture system. For the in vivo cell tracking, DBCO-650-labeled chondrocytes (1 × 10(6) cells) seeded on the 3D scaffold were subcutaneously implanted into mice and the transplanted DBCO-650-labeled chondrocytes could be effectively tracked in the prolonged time period of 4 weeks using NIRF imaging technology. Furthermore, this new cell labeling and tracking technology had minimal effect on cartilage formation in vivo.

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