Abstract

In spite of recognized limitations in capturing species-specific responses and high costs, rodent models remain commonly used in prostate cancer metastasis research, due largely to the lack of available alternatives. We aim to develop an in vitro culture system to study prostate cancer response to a simulated bone microenvironment, which may be used to understand early events in prostate metastasis to bone or for drug screening applications. To achieve this, mesenchymal stem cells and endothelial cells were isolated and cocultured to form a vascularized bone analogue. Endothelial cells were found to exert osteopotentiating effects on mesenchymal stem cell differentiation, and reciprocal effects by the stromal cells were found to stimulate vasculogenic responses, suggesting the possible utility of this system to elicit three-way interactions between endothelial, mesenchymal, and prostate cancer cells. We further developed the use of fluorescently labeled cells which could be used to concurrently track cellular migration, proliferation, and morphometric analysis. We demonstrate the concurrent, real-time visualization of prostate cancer and endothelial cells, which may be useful for evaluation of spatiotemporal changes at a single-cell level. When prostate cancer cell proliferation on various substrates was measured, it was found that the use of coculture systems may provide a better reflection of conditions in vivo, highlighting the potential utility as a model system.

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