Optimization of Aqueous Biphasic Tumor Spheroid Microtechnology for Anti-cancer Drug Testing in 3D Culture
Open accessAbstract
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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Spheroid cell culture has been developed for research on cancer, stem cells, and many organs, including liver, kidney, and heart.
“This approach has been developed in the research of cancer (7, 76, 102), stem cells (17, 120), and many organs (such as liver, kidney, heart, etc.) (3,16,64,75).”
From Modeling physiological events in 2D vs. 3D cell culture (2017) · cites this paper as [76]
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- Authors
- Lemmo, S., Atefi, E., Luker, G. D., & Tavana, H.
- Affiliations
- University of Akron (US)
- University of Michigan (US)
- Corresponding author
- Tavana, Hossein
- Venue
- Cellular and Molecular Bioengineering
- Published
- 2014-08-05
- Volume
- 7
- Issue
- 3
- Pages
- 344-354
- ISSN
- 1865-5025 (ISSN-L)
- 1865-5033
- PMC
- PMC4159185
- OpenAlex
- W1985471235
- Cited by
- 71
- References
- 49
- Citations by year
- 2025: 6 · 2024: 2 · 2023: 8 · 2022: 4 · 2021: 5 · 2020: 9 · 2019: 7 · 2018: 6 · 2017: 9 · 2016: 9 · 2015: 6
- FWCI
- 4.15
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Cellular and Molecular Bioengineering (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Funders
- National Institutes of Health
- National Cancer Institute (R21CA182333)
- Topics
- Physical Sciences › Engineering › Biomedical Engineering › 3D Printing in Biomedical Research
- Health Sciences › Medicine › Oncology › Cancer Cells and Metastasis
- Physical Sciences › Chemical Engineering › Filtration and Separation › Chemical and Physical Properties in Aqueous Solutions
- Keywords
- tumor spheroids, aqueous two-phase systems, anticancer drug screening, 3D cell culture, microwells, polyethylene glycol, liquid handling robot, cisplatin, paclitaxel resistance, high-throughput drug screening
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