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Optimization of Aqueous Biphasic Tumor Spheroid Microtechnology for Anti-cancer Drug Testing in 3D Culture

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Abstract

Tumor spheroids are three-dimensional clusters of cancer cells that exhibit characteristics of poorly perfused tumors and hence present a relevant model for testing the efficacy of anti-cancer compounds. The use of spheroids for drug screening is hindered by technological complexities for high throughput generation of consistent size spheroids individually addressable by drug compounds. Here we present and optimize a simple spheroid technology based on the use of an aqueous two-phase system. Cancer cells confined in a drop of the denser aqueous dextran phase are robotically dispensed into a microwell containing the immersion aqueous polyethylene glycol phase. Cells remain within the drop and form a viable spheroid, without a need for any external stimuli. The size of resulting spheroids is sensitive to volume variations of dispensed drops from the air displacement pipetting head of a commercial liquid handling robot. Therefore, we parametrically optimize the process of dispensing of dextran phase drops. For a given cell density, this optimization reproducibly generates consistent size spheroids in standard 96-well plates. In addition, we evaluate the use of a commercial biochemical assay to examine cellular viability of cancer cell spheroids. Spheroids show a dose-dependent response to cisplatin similar to a monolayer culture. However unlike their two-dimensional counterpart, spheroids exhibit resistance to paclitaxel treatment. This technology, which uses only commercially-available reagents and equipment, can potentially expedite anti-cancer drug discovery. Although the use of robotics makes the ATPS spheroid technology particularly useful for drug screening applications, this approach is compatible with simpler liquid handling techniques such as manual micropipetting and offers a straightforward method of 3D cell culture in research laboratories.

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Abbreviations

ATPS:

Aqueous two-phase system

3D:

Three-dimensional

PEG:

Polyethylene glycol

DEX:

Dextran

CCS:

Cancer cell spheroids

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Acknowledgements

This work is supported by a Grant from the National Institutes of Health R21CA182333.

Conflict of interest

Stephanie Lemmo, Ehsan Atefi, Gary D. Luker, and Hossein Tavana declare that they have no conflict of interest.

Ethical Standards

This work involved no human subjects or animal studies.

Author information

Authors and Affiliations

  1. Department of Biomedical Engineering, The University of Akron, Akron, OH, 44325, USA

    Stephanie Lemmo, Ehsan Atefi & Hossein Tavana

  2. Department of Radiology, University of Michigan, Ann Arbor, MI, 48109, USA

    Gary D. Luker

  3. Department of Microbiology and Immunology, University of Michigan, Ann Arbor, MI, 48109, USA

    Gary D. Luker

  4. Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI, 48109, USA

    Gary D. Luker

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  1. Stephanie Lemmo
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  2. Ehsan Atefi
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Corresponding author

Correspondence to Hossein Tavana.

Additional information

Associate Editor Cynthia A. Reinhart-King oversaw the review of this article.

This paper is part of the 2014 Young Innovators Issue.

This paper is part of the 2014 Young Innovators Issue.

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Lemmo, S., Atefi, E., Luker, G.D. et al. Optimization of Aqueous Biphasic Tumor Spheroid Microtechnology for Anti-cancer Drug Testing in 3D Culture. Cel. Mol. Bioeng. 7, 344–354 (2014). https://doi.org/10.1007/s12195-014-0349-4

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