Magnetic force-based cell patterning for evaluation of the effect of stromal fibroblasts on invasive capacity in 3Dcultures

Biomimetic cell culture systems that recreate tumor microenvironments are necessary in understanding the progression of cancer cells in cell-to-cell interaction and in cell-to-extracellular matrix interaction. We have developed a three-dimensional spheroid array embedded in collagen for evaluation of the effect of stromal fibroblasts associated with cancer cells. When the breast epithelial cancer cell model MCF10A/myr-Akt1 was magnetically labeled and aligned in the array by an external magnetic force using a pin-holder device and a magnet, a stellate configuration was observed. Changes in MCF10A/myr-Akt1 cell behavior were only slight when normal human dermal fibroblasts (NHDF) cells coexisted in collagen (indirect-interaction array). In contrast, when NHDF were magnetically labeled and patterned together with MCF10A/myr-Akt1 (direct-interaction array), spreading and progression were observed along with NHDF. Cell image analysis indicated that the length and area were statistically significantly increased in the direct-interaction array compared to the MCF10A/myr-Akt1 alone or to the indirect-interaction array. A cell susceptibility assay was undertaken with breast cancer MDA-MB-231 associated with NHDF in the indirect-interaction array. Interestingly, although distinct suppression of cell movement and proliferation was observed with 100 μM of collagenase inhibitor, formation of invadepodia significantly increased with coexistent NHDF. Since cancer progression is influenced by its microenvironment, this magnetic cell-patterning method which clarifies direct and indirect effects of stromal cells on invasion and proliferation, is well suited for evaluation and design of more efficient approaches in cancer prevention and treatment.

► The 3D tumor spheroid cell array in ECM was developed by magnetic patterning.
► Direct and indirect interaction of fibroblasts on tumor cell invasion was evaluated.
► Fibroblasts enhanced tumor cell aggressiveness with anticancer agent treatment.