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

Title
Bioorthogonal Copper Free Click Chemistry for Labeling and Tracking of Chondrocytes In Vivo
Authors
권익찬김광명류주희이상민장혜윤윤화인이지영나진희이혁진강선웅이슬기조영우
Keywords
Bioorthogonal Click Chemistry; DBCO-650; Cell therapy; Chondrocytes
Issue Date
2016-04
Publisher
Bioconjugate chemistry
Citation
VOL 27, NO 4-936
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 (Ac(4)ManNAz) to generate unnatural azide groups (-N-3) 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 x 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.
URI
http://pubs.kist.re.kr/handle/201004/64962
ISSN
1043-1802
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KIST Publication > Article
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