Continuous, real-time bioimaging of chemical bioavailability and toxicology using autonomously bioluminescent human cell lines

Tingting Xu; Dan M. Close; James D. Webb; Sarah L. Price; Steven A. Ripp; Gary S. Sayler

doi:10.1117/12.2015030

29 May 2013 Continuous, real-time bioimaging of chemical bioavailability and toxicology using autonomously bioluminescent human cell lines

Tingting Xu, Dan M. Close, James D. Webb, Sarah L. Price, Steven A. Ripp, Gary S. Sayler

Author Affiliations +

Proceedings Volume 8723, Sensing Technologies for Global Health, Military Medicine, and Environmental Monitoring III; 872310 (2013) https://doi.org/10.1117/12.2015030
Event: SPIE Defense, Security, and Sensing, 2013, Baltimore, Maryland, United States

Abstract

Bioluminescent imaging is an emerging biomedical surveillance strategy that uses external cameras to detect in vivo light generated in small animal models of human physiology or in vitro light generated in tissue culture or tissue scaffold mimics of human anatomy. The most widely utilized of reporters is the firefly luciferase (luc) gene; however, it generates light only upon addition of a chemical substrate, thus only generating intermittent single time point data snapshots. To overcome this disadvantage, we have demonstrated substrate-independent bioluminescent imaging using an optimized bacterial bioluminescence (lux) system. The lux reporter produces bioluminescence autonomously using components found naturally within the cell, thereby allowing imaging to occur continuously and in real-time over the lifetime of the host. We have validated this technology in human cells with demonstrated chemical toxicological profiling against exotoxin exposures at signal strengths comparable to existing luc systems (~1.33 × 10⁷ photons/second). As a proof-in-principle demonstration, we have engineered breast carcinoma cells to express bioluminescence for real-time screening of endocrine disrupting chemicals and validated detection of 17β-estradiol (EC₅₀ = ~ 10 pM). These and other applications of this new reporter technology will be discussed as potential new pathways towards improved models of target chemical bioavailability, toxicology, efficacy, and human safety.

Citation Download Citation

Tingting Xu, Dan M. Close, James D. Webb, Sarah L. Price, Steven A. Ripp, and Gary S. Sayler "Continuous, real-time bioimaging of chemical bioavailability and toxicology using autonomously bioluminescent human cell lines", Proc. SPIE 8723, Sensing Technologies for Global Health, Military Medicine, and Environmental Monitoring III, 872310 (29 May 2013); https://doi.org/10.1117/12.2015030

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