Fluorescent microparticles for sensing cell microenvironment oxygen levels within 3D scaffolds

Oxygen diffusion through tissues is a critical factor in maintaining physiologic cell processes and growing engineered tissues in vitro. We have recently reported a novel fluorescent microparticle system for sensing cell microenvironment oxygen levels within 3D scaffolds. Cell response to suboptimal oxygen concentration is controlled by the hypoxia inducible factor-1 (HIF-1). Thus, HIF-1 is an ideal marker for investigating cellular response to oxygen levels in engineered tissues. We have created a cell line consisting of human C8161 subcutaneous melanoma cells transfected to express an EGFP-HIF-1α fusion. Expression of this fusion protein may be quantified with analysis tools such as microscopy or flow-cytometry. In this study, we present the preliminary characterization of these cells and we demonstrate that they are capable of expressing measurable quantities of EGFP-HIF-1α fusion protein in response to prolonged exposure to hypoxia.