title: Bioorthogonal Copper Free Click Chemistry for Labeling and Tracking of Chondrocytes In Vivo.
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.
authors.affiliation: Center for Theragnosis, Biomedical Research Institute, Korea Institute of Science and Technology , Hwarangno 14-gil 6, Seongbuk-gu, Seoul 136-791, Republic of Korea.|Departments of Chemical Engineering and Bionanotechnology, Hanyang University , Ansan, Gyeonggi-do 426-791, Republic of Korea.
authors.forename: Hwa In
authors.initials: HI
authors.lastname: Yoon
authors.affiliation: College of Pharmacy, Graduate School of Pharmaceutical Sciences, Ewha Womans University , Seoul 120-750, Republic of Korea.
authors.forename: Ji Young
authors.initials: JY
authors.lastname: Yhee
authors.affiliation: The Russell H. Morgan Department of Radiology and Radiological Sciences, Johns Hopkins University School of Medicine , Baltimore, Maryland United States.
authors.forename: Jin Hee
authors.initials: JH
authors.lastname: Na
authors.affiliation: The Russell H. Morgan Department of Radiology and Radiological Sciences, Johns Hopkins University School of Medicine , Baltimore, Maryland United States.
authors.forename: Sangmin
authors.initials: S
authors.lastname: Lee
authors.affiliation: College of Pharmacy, Graduate School of Pharmaceutical Sciences, Ewha Womans University , Seoul 120-750, Republic of Korea.
authors.forename: Hyukjin
authors.initials: H
authors.lastname: Lee
authors.affiliation: Next-generation Pharmaceutical Research Center, Korea Institute of Toxicology , Daejeon 305-343, Republic of Korea.|Human and Environmental Toxicology Program, University of Science and Technology (UST) , Daejeon 305-350, Republic of Korea.
authors.forename: Sun-Woong
authors.initials: SW
authors.lastname: Kang
authors.affiliation: Center for Theragnosis, Biomedical Research Institute, Korea Institute of Science and Technology , Hwarangno 14-gil 6, Seongbuk-gu, Seoul 136-791, Republic of Korea.|Department of Biomedical Engineering, University of Science and Technology (UST) , Seoul 136-791, Republic of Korea.
authors.forename: Hyeyoun
authors.initials: H
authors.lastname: Chang
authors.affiliation: Center for Theragnosis, Biomedical Research Institute, Korea Institute of Science and Technology , Hwarangno 14-gil 6, Seongbuk-gu, Seoul 136-791, Republic of Korea.
authors.forename: Ju Hee
authors.initials: JH
authors.lastname: Ryu
authors.affiliation: The Russell H. Morgan Department of Radiology and Radiological Sciences, Johns Hopkins University School of Medicine , Baltimore, Maryland United States.
authors.forename: Seulki
authors.initials: S
authors.lastname: Lee
authors.affiliation: Center for Theragnosis, Biomedical Research Institute, Korea Institute of Science and Technology , Hwarangno 14-gil 6, Seongbuk-gu, Seoul 136-791, Republic of Korea.
authors.forename: Ick Chan
authors.initials: IC
authors.lastname: Kwon
authors.affiliation: Departments of Chemical Engineering and Bionanotechnology, Hanyang University , Ansan, Gyeonggi-do 426-791, Republic of Korea.
authors.forename: Yong Woo
authors.initials: YW
authors.lastname: Cho
authors.affiliation: Center for Theragnosis, Biomedical Research Institute, Korea Institute of Science and Technology , Hwarangno 14-gil 6, Seongbuk-gu, Seoul 136-791, Republic of Korea.|Department of Biomedical Engineering, University of Science and Technology (UST) , Seoul 136-791, Republic of Korea.
authors.forename: Kwangmeyung
authors.initials: K
authors.lastname: Kim
mesh_terms: D000818:Animals; D002356:Cartilage / Q000166:cytology*; D019902:Chondrocytes / Q000166:cytology*; D057930:Click Chemistry; D003300:Copper / Q000737:chemistry*; D005434:Flow Cytometry; D051379:Mice; D023822:Tissue Engineering
chemical_list: D003300:Copper