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Development of High-Throughput Quantitative Assays for Glucose Uptake in Cancer Cell Lines



Metabolism, and especially glucose uptake, is a key quantitative cell trait that is closely linked to cancer initiation and progression. Therefore, developing high-throughput assays for measuring glucose uptake in cancer cells would be enviable for simultaneous comparisons of multiple cell lines and microenvironmental conditions. This study was designed with two specific aims in mind: the first was to develop and validate a high-throughput screening method for quantitative assessment of glucose uptake in “normal” and tumor cells using the fluorescent 2-deoxyglucose analog 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxyglucose (2-NBDG), and the second was to develop an image-based, quantitative, single-cell assay for measuring glucose uptake using the same probe to dissect the full spectrum of metabolic variability within populations of tumor cells in vitro in higher resolution.


The kinetics of population-based glucose uptake was evaluated for MCF10A mammary epithelial and CA1d breast cancer cell lines, using 2-NBDG and a fluorometric microplate reader. Glucose uptake for the same cell lines was also examined at the single-cell level using high-content automated microscopy coupled with semi-automated cell-cytometric image analysis approaches. Statistical treatments were also implemented to analyze intra-population variability.


Our results demonstrate that the high-throughput fluorometric assay using 2-NBDG is a reliable method to assess population-level kinetics of glucose uptake in cell lines in vitro. Similarly, single-cell image-based assays and analyses of 2-NBDG fluorescence proved an effective and accurate means for assessing glucose uptake, which revealed that breast tumor cell lines display intra-population variability that is modulated by growth conditions.


These studies indicate that 2-NBDG can be used to aid in the high-throughput analysis of the influence of chemotherapeutics on glucose uptake in cancer cells.

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DMEM without serum and supplements






Arbitrary fluorescence units


Confidence interval


Epidermal growth factor


2-Deoxy2-[F-18] fluoro-d-glucose


Fetal bovine serum


Glucose transporters


High-content automated microscopy


Phosphate-buffered saline


Positron emission tomography


Standard error of the mean


DMEM with serum and supplements


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We would like to acknowledge the following funding source for support of this work: NCI U54-CA113007 awarded to VQ. We would also like to thank Dr. David Degraff for his help with the revising the scientific content of this manuscript and for his insightful comments and critiques.

Conflict of Interest Statement

The authors declare that they have no conflict of interest.

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Correspondence to Vito Quaranta.

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Hassanein, M., Weidow, B., Koehler, E. et al. Development of High-Throughput Quantitative Assays for Glucose Uptake in Cancer Cell Lines. Mol Imaging Biol 13, 840–852 (2011). https://doi.org/10.1007/s11307-010-0399-5

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Key words

  • 2-NBDG
  • Metabolism
  • High-throughput
  • Single-cell
  • Glycolysis