Cell labeling and tracking method without distorted signals by phagocytosis of macrophages.
Cell labeling and tracking method without distorted signals by phagocytosis of macrophages.
复制标题
作者：
Kang SW;Lee S;Na JH;Yoon HI;Lee DE;Koo H;Cho YW;Kim SH;Jeong SY;Kwon IC;Choi K;Kim K
细胞标记和追踪是了解体内接种细胞的生物学机制和治疗效果的重要过程。已经进行了许多尝试来标记和跟踪体内接种的细胞;然而，这些方法由于其生物学效应而具有局限性，包括巨噬细胞的二次吞噬和遗传修饰。在这里，我们研究了一种新的细胞标记和跟踪策略的基础上代谢糖工程和生物正交点击化学。我们首先用四乙酰化的N-叠氮乙酰基-D-甘露糖胺处理细胞，以在靶细胞表面上产生具有叠氮基的非天然唾液酸。然后将叠氮化物标记的细胞移植到小鼠肝脏中，并将二苄基环辛炔缀合的Cy 5（DBCO-Cy 5）静脉内注射到小鼠中，以与体内靶细胞表面上的叠氮化物基团化学结合，用于靶细胞可视化。利用糖工程技术可以在靶细胞表面人工诱导产生非天然的含叠氮基的唾液酸。然后，我们使用生物正交点击化学通过DBCO-Cy 5在体内追踪细胞表面上的叠氮基团。重要的是，标记功效增强，巨噬细胞吞噬作用的假信号减少。这种策略对于细胞标记和跟踪将是非常有用的。
Cell labeling and tracking are important processes in understanding biologic mechanisms and the therapeutic effect of inoculated cells in vivo. Numerous attempts have been made to label and track inoculated cells in vivo; however, these methods have limitations as a result of their biological effects, including secondary phagocytosis of macrophages and genetic modification. Here, we investigated a new cell labeling and tracking strategy based on metabolic glycoengineering and bioorthogonal click chemistry. We first treated cells with tetra-acetylated N-azidoacetyl-D-mannosamine to generate unnatural sialic acids with azide groups on the surface of the target cells. The azide-labeled cells were then transplanted to mouse liver, and dibenzyl cyclooctyne-conjugated Cy5 (DBCO-Cy5) was intravenously injected into mice to chemically bind with the azide groups on the surface of the target cells in vivo for target cell visualization. Unnatural sialic acids with azide groups could be artificially induced on the surface of target cells by glycoengineering. We then tracked the azide groups on the surface of the cells by DBCO-Cy5 in vivo using bioorthogonal click chemistry. Importantly, labeling efficacy was enhanced and false signals by phagocytosis of macrophages were reduced. This strategy will be highly useful for cell labeling and tracking.
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DOI：
10.1084/jem.20061916
发表时间：
2007-02-19
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Behfar A;Perez-Terzic C;Faustino RS;Arrell DK;Hodgson DM;Yamada S;Puceat M;Niederländer N;Alekseev AE;Zingman LV;Terzic A
通讯作者：
Terzic A
影响因子：
4
作者：
Dehnert, Karen W.;Beahm, Brendan J.;Huynh, Thinh T.;Baskin, Jeremy M.;Laughlin, Scott T.;Wang, Wei;Wu, Peng;Amacher, Sharon L.;Bertozzi, Carolyn R.
通讯作者：
Bertozzi, Carolyn R.
影响因子：
4.7
作者：
Hong, Vu;Steinmetz, Nicole F.;Manchester, Marianne;Finn, M. G.
通讯作者：
Finn, M. G.
影响因子：
16.6
作者：
Ning, Xinghai;Guo, Jun;Boons, Geert-Jan
通讯作者：
Boons, Geert-Jan
影响因子：
10.8
作者：
Kim, Betty Y. S.;Jiang, Wen;Chan, Warren C. W.
通讯作者：
Chan, Warren C. W.