Embedded multicellular spheroids as a biomimetic 3D cancer model for evaluating drug and drug-device combinations - PubMed (original) (raw)

Embedded multicellular spheroids as a biomimetic 3D cancer model for evaluating drug and drug-device combinations

Kristie M Charoen et al. Biomaterials. 2014 Feb.

Abstract

Multicellular aggregates of cells, termed spheroids, are of interest for studying tumor behavior and for evaluating the response of pharmacologically active agents. Spheroids more faithfully reproduce the tumor macrostructure found in vivo compared to classical 2D monolayers. We present a method for embedding spheroids within collagen gels followed by quantitative and qualitative whole spheroid and single cell analyses enabling characterization over the length scales from molecular to macroscopic. Spheroid producing and embedding capabilities are demonstrated for U2OS and MDA-MB-231 cell lines, of osteosarcoma and breast adenocarcinoma origin, respectively. Finally, using the MDA-MB-231 tumor model, the chemotherapeutic response between paclitaxel delivery as a bolus dose, as practiced in the clinic, is compared to delivery within an expansile nanoparticle. The expansile nanoparticle delivery route provides a superior outcome and the results mirror those observed in a murine xenograft model. These findings highlight the synergistic beneficial results that may arise from the use of a drug delivery system, and the need to evaluate both drug candidates and delivery systems in the research and preclinical screening phases of a new cancer therapy development program.

Keywords: 3D cell culture; Cancer model; Cell migration; Collagen; Drug delivery; Spheroid; Tumor mimic.

Copyright © 2013 Elsevier Ltd. All rights reserved.

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Figures

Figure 1

Figure 1

Creation of Embedded Spheroids: Spheroid formation is encouraged by placing a suspension of cells (red) in media (pink) on agarose (yellow) coated wells. After 72 hours, a spheroid is formed, and then transferred into a collagen gel.

Figure 2

Figure 2

Controllable Spheroid Size: Spheroid size can be controlled by varying cell seeding number. Sizing based on cell seeding number was demonstrated with both bone (top) and breast (bottom) cancer cells.

Figure 2

Figure 2

Controllable Spheroid Size: Spheroid size can be controlled by varying cell seeding number. Sizing based on cell seeding number was demonstrated with both bone (top) and breast (bottom) cancer cells.

Figure 3

Figure 3

U2OS Spheroid Growth in Collagen: Bone cancer spheroids grown in collagen gels with varying mechanical properties demonstrated the most growth in 3-4 mg/mL, growing less in both stiffer and weaker gels.

Figure 3

Figure 3

U2OS Spheroid Growth in Collagen: Bone cancer spheroids grown in collagen gels with varying mechanical properties demonstrated the most growth in 3-4 mg/mL, growing less in both stiffer and weaker gels.

Figure 4

Figure 4

MDA-MB 231 Spheroid Growth in Collagen: Breast cancer spheroids grown in collagen gels with varying mechanical properties demonstrated the most growth in the weakest collagen gels.

Figure 4

Figure 4

MDA-MB 231 Spheroid Growth in Collagen: Breast cancer spheroids grown in collagen gels with varying mechanical properties demonstrated the most growth in the weakest collagen gels.

Figure 5

Figure 5

Pattern of Metabolic Activity: A spheroid in collagen was stained with a calcein based stain (green) to show metabolically active cells and an ethidium based stain (red) to demonstrate cells with a compromised membrane. The red staining on the interior indicates a mostly dead core, whereas the green cells shows a metabolically active outer ring of cells. The inset quantitatively demonstrates that 85.5% of the spheroid is alive after 3 days via FACS.

Figure 6

Figure 6

Metabolic Activity of Spheroids: Disaggregated spheroids were plated in a monolayer and the metabolic activity was compared to cells grown only in a monolayer. Although there were approximately 60,000 cells, the overall metabolic level is comparable to 30,000 cells.

Figure 7

Figure 7

Nanoparticle Penetration: Within 24 hours fluorescently labeled nanoparticles were able to fully penetrate a 20,000 cell MDA-MB 231 spheroid. Scale bar is 100 _μ_m.

Figure 8

Figure 8

Spheroid Treatment with Nanoparticles: Paclitaxel, a chemotherapeutic drug was delivered via bolus dose or nanoparticles for 24 hours before the treatment was removed. The nanoparticle delivery method was the most effective at reducing spheroid size.

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