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


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 17?-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.

SUBMITTER: Xu T 

PROVIDER: S-EPMC4621765 | biostudies-literature | 2013 May

REPOSITORIES: biostudies-literature

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

Xu Tingting T   Close Dan M DM   Webb James D JD   Price Sarah L SL   Ripp Steven A SA   Sayler Gary S GS  

Proceedings of SPIE--the International Society for Optical Engineering 20130501


Bioluminescent imaging is an emerging biomedical surveillance strategy that uses external cameras to detect <i>in vivo</i> light generated in small animal models of human physiology or <i>in vitro</i> light generated in tissue culture or tissue scaffold mimics of human anatomy. The most widely utilized of reporters is the firefly luciferase (<i>luc</i>) gene; however, it generates light only upon addition of a chemical substrate, thus only generating intermittent single time point data snapshots  ...[more]

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