Physiological Effects of Ac4ManNAz and Optimization of Metabolic Labeling for Cell Tracking

Abstract

Metabolic labeling techniques are powerful tools for cell labeling, tracking and proteomic analysis. However, at present, the effects of the metabolic labeling agents on cell metabolism and physiology are not known. To address this question, in this study, we analyzed the effects of cells treated with Ac4ManNAz through microarray analysis and analyses of membrane channel activity, individual bio-physiological properties, and glycolytic flux. According to the results, treatment with 50 μM Ac4ManNAz led to the reduction of major cellular functions, including energy generation capacity, cellular infiltration ability and channel activity. Interestingly, 10 μM Ac4ManNAz showed the least effect on cellular systems and had a sufficient labeling efficiency for cell labeling, tracking and proteomic analysis. Based on our results, we suggest 10 μM as the optimum concentration of Ac4ManNAz for in vivo cell labeling and tracking. Additionally, we expect that our approach could be used for cell-based therapy for monitoring the efficacy of molecule delivery and the fate of recipient cells.

Exploring the Potential of β-Catenin O-GlcNAcylation by Using Fluorescence-Based Engineering and Imaging

“…As shown in Figure 4 b, no modification of the morphology of the cells was observed even at the highest dose of 200 µM of the chemical reporter, showing that the toxicity of the azidosugar concentration was negligible even at the highest concentrations. Acetic acid release inside cells upon enzymatic deacetylation of Ac 4 GalNAz did not seem to induce cytotoxicity, as was previously reported for Ac 4 ManNAz (>50 μM) [ 23 ]. In order to promote high FRET efficiencies, we chose the highest non-toxic Ac 4 GlcNAz concentration of 200 µM.…”

Effect of Cell Labeling on the Function of Human Pluripotent Stem Cell-Derived Cardiomyocytes

“…Many hurdles remain to the widespread acceptance of labeling for cell tracking and labeling for stem cell-derived cardiomyocytes because the functional changes of labeling agents in cells are not fully understood. Although these materials have biocompatibility in the cells, many previous studies have shown that the carbocyanine dyes with highly lipophilic nature and nano-materials such as QD specifically led to changes in bio-physiological function, cellular signaling pathways and interference with cell-cell communication ( 4 - 6 , 25 , 26 ). Indeed, carbocyanine dyes showed at high concentrations in cancer cell lines ( 6 ).…”

“…The transportation of labeling agents into the cells could change the biological responses of cells, such as metabolism, proliferation, and migration ( 4 - 6 ). Labeling agent-rich regions cause changes in the physical properties of the cell surface and thereby determine cellular characteristics, i.e., the cell surface, repulsion between cells, subsequent migration, and the uptake of ion ( 7 - 9 ).…”

Facile metabolic glycan labeling strategy for exosome tracking

“…It is not clear how does this occur, but a similar trend has been observed in literatures. [26, 30, 31]…”