Aqueous biphasic cancer cell migration assay enables robust, high‐throughput screening of anti‐cancer compounds

Lemmo, Stephanie; Nasrollahi, Samila; Tavana, Hossein

doi:10.1002/biot.201300227

Cited by 17 publications

(20 citation statements)

References 38 publications

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“…Therefore, ATPS is solely used as a patterning medium to generate spheroids. Micropatterning with ATPS has additionally facilitated the generation of cancer cell niches for migration studies …”

Section: Resultsmentioning

confidence: 99%

Engineered Breast Cancer Cell Spheroids Reproduce Biologic Properties of Solid Tumors

Ham

Joshi

Luker

et al. 2016

Adv Healthcare Materials

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Solid tumors develop as three-dimensional tissue constructs. As tumors grow larger, spatial gradients of nutrients and oxygen and inadequate diffusive supply to cells distant from vasculature develops. Hypoxia initiates signaling and transcriptional alterations to promote survival of cancer cells and generation of cancer stem cells (CSCs) that have self-renewal and tumor-initiation capabilities. Both hypoxia and CSCs are associated with resistance to therapies and tumor relapse. Here, we conduct a systematic study to demonstrate that 3D cancer cell models, known as tumor spheroids, generated with a polymeric aqueous two-phase system (ATPS) technology capture these important biological processes. Similar to solid tumors, spheroids of triple negative breast cancer (TNBC) cells deposit major extracellular matrix proteins. Our molecular analysis establishes presence of hypoxic cells in the core region and expression of CSC gene and protein markers including CD24, CD133, and Nanog. Importantly, these spheroids resist treatment with chemotherapy drugs. We successfully target drug resistant spheroids through a combination treatment approach using a hypoxia-activated prodrug, TH-302, and a chemotherapy drug, doxorubicin. This study demonstrates that ATPS spheroids recapitulate important biological and functional properties of solid tumors and provide a unique model for studies in cancer research.

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Section: Resultsmentioning

confidence: 99%

Engineered Breast Cancer Cell Spheroids Reproduce Biologic Properties of Solid Tumors

Ham

Joshi

Luker

et al. 2016

Adv Healthcare Materials

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“…Drop diameter data from experiments at air volumes of 0.3–1.5 μl follow a linear trend, indicating that data are normally distributed. (30, 31) Different slopes of the lines indicate differences between standard deviations of data from these air volumes. (32) We confirmed that data obtained for drop diameter by varying each of the three parameters within the pre-defined range of Fig.…”

Section: Resultsmentioning

confidence: 99%

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

et al. 2014

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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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“…After 12 hours of incubation, the ATPS was replaced with growth medium only (control) or medium containing a test compound. The area of the cell-excluded gap within well plates averaged to 3.33 ± 0.09 mm 2 27. …”

Section: Methodsmentioning

confidence: 99%

“…We have previously evaluated this robotic technology for high throughput compound screening against cell migration and shown that it consistently returns a Z ′ factor of greater than 0.7, demonstrating its quality and robust performance. 27 Using this migration technology, our screening identified several natural compounds that effectively blocked motility of cells. Subsequent molecular analysis showed that phytochemicals interfered with dysregulated kinase signaling and scavenged excessive intracellular free radicals in TNBC cells.…”

Section: Introductionmentioning

confidence: 99%

Phytochemicals potently inhibit migration of metastatic breast cancer cells

et al. 2015

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Cell migration is a major process that drives metastatic progression of cancers, the major cause of cancer death. Existing chemotherapeutic drugs have limited efficacy to prevent and/or treat metastasis, emphasizing the need for new treatments. We focus on triple negative breast cancer (TNBC), the subtype of breast cancer with worst prognosis and no standard chemotherapy protocols. Here we demonstrate that a group of natural compounds, known as phytochemicals, effectively block migration of metastatic TNBC cells. Using a novel cell micropatterning technology, we generate consistent migration niches in standard 96-well plates where each well contains a cell-excluded gap within a uniform monolayer of cells. Over time, cells migrate into and occupy the gap. Treating TNBC cells with non-toxic concentrations of phytochemicals significantly blocks motility of cells. Using a molecular analysis approach, we show that anti-migratory property of phytochemicals is partly due to their inhibitory effects on phosphorylation of ERK1/2. This study provides a framework for future studies to understand molecular targets of phytochemicals and evaluate their effectiveness in inhibiting metastasis in animal models of cancer.

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Aqueous biphasic cancer cell migration assay enables robust, high‐throughput screening of anti‐cancer compounds

Cited by 17 publications

References 38 publications

Engineered Breast Cancer Cell Spheroids Reproduce Biologic Properties of Solid Tumors

Engineered Breast Cancer Cell Spheroids Reproduce Biologic Properties of Solid Tumors

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

Phytochemicals potently inhibit migration of metastatic breast cancer cells

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