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

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• 作者：Hwa In Yoon ; Ji Young Yhee ; Jin Hee Na ; Sangmin Lee ; Hyukjin Lee ; Sun-Woong Kang ; Hyeyoun Chang ; Ju Hee Ryu ; Seulki Lee ; Ick Chan Kwon ; Yong Woo Cho ; Kwangmeyung Kim
• 刊名：Bioconjugate Chemistry
• 出版年：2016
• 出版时间：April 20, 2016
• 年：2016
• 卷：27
• 期：4
• 页码：927-936
• 全文大小：666K
• 年卷期：0
• ISSN：1520-4812

文摘

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<sub>4sub>ManNAz) to generate unnatural azide groups (-N<sub>3sub>) 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 Ac<sub>4sub>ManNAz and DBCO-650 to the cell culture system. For the in vivo cell tracking, DBCO-650-labeled chondrocytes (1 × 10<sup>6sup> 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.