A Flow Cytometric Clonogenic Assay Reveals the Single-Cell Potency of Doxorubicin
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Hammond_A flow cytometric.pdf
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Author(s)
Kulkarni, Chethana
Betts, Alison M.
Maass, Katie F.
Quadir, Mohiuddin Abdul
Hammond, Paula T.
Wittrup, Karl Dane
Date Issued
January 2016
Journal
Journal of Pharmaceutical Sciences
Publisher
Wiley Blackwell
Citation
Maass, Katie F. et al. “A Flow Cytometric Clonogenic Assay Reveals the Single-Cell Potency of Doxorubicin.” Journal of Pharmaceutical Sciences 104.12 (2015): 4409–4416.
Version
Author's final manuscript
Abstract
Standard cell proliferation assays use bulk media drug concentration to ascertain the potency of chemotherapeutic drugs; however, the relevant quantity is clearly the amount of drug actually taken up by the cell. To address this discrepancy, we have developed a flow cytometric clonogenic assay to correlate the amount of drug in a single cell with the cell’s ability to proliferate using a cell tracing dye and doxorubicin, a naturally fluorescent chemotherapeutic drug. By varying doxorubicin concentration in the media, length of treatment time, and treatment with verapamil, an efflux pump inhibitor, we introduced 10[superscript 5]–10[superscript 10] doxorubicin molecules per cell; then used a dye-dilution assay to simultaneously assess the number of cell divisions. We find that a cell’s ability to proliferate is a surprisingly conserved function of the number of intracellular doxorubicin molecules, resulting in single-cell IC[subscript 50] values of 4–12 million intracellular doxorubicin molecules. The developed assay is a straightforward method for understanding a drug’s single-cell potency and can be used for any fluorescent or fluorescently labeled drug, including nanoparticles or antibody–drug conjugates.
MIT Department
Massachusetts Institute of Technology. Department of Biological Engineering
Massachusetts Institute of Technology. Department of Chemical Engineering
Koch Institute for Integrative Cancer Research at MIT
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DOI of Published Version
https://doi.org/10.1002/jps.24631
