From a regulatory and research perspective, the outcomes from in vitro toxicology testing of foods, dietary supplements, and additives can help stratify which compounds merit further safety assessment using analytical and/or classical in vivo toxicology methodology. In vitro toxicology methods carry the advantages of yielding informative data about the potential chemical hazards in foods as well as their cellular mechanisms of action in a relatively short timeframe than would be obtained from an in vivo study. The fact that the majority of in vitro toxicology screening methods are commercially sourced, however, ultimately results in the use of proprietary reagents that (a) are expensive and (b) can impede troubleshooting efforts in some situations. Moreover, the need to screen multiple food-related ingredients in parallel at multiple concentrations and durations of exposure creates the requirement for more efficient in vitro toxicity screening tools. To help address some of these issues, we have developed a ‘designer’ human kidney cell line we coined HK2-Vi. This cell line was genetically engineered by lentivirus to stably encode the Perceval HR biosensor, which differentially fluoresces when bound to ATP vs. ADP. The ratio of ATP to ADP indicates the proportion of cells that are alive. We designed this novel in vitro tool to be an inexpensive kinetic assay that yields data on short- or long-term kidney cell viability after toxin exposure. We tailored this prototypic tool towards kidney cell testing because the kidneys are inherently vulnerable to the effects of toxins, due to their important role in concentrating blood solutes within their specialized compartments. Proof-of-principle experiments using food additive or metabolic poison compounds will be followed by comprehensive testing to help establish HK2-Vi as a rapid and cost-efficient tool for in vitro toxicology screening of foods, dietary supplements, and additives.

Novel tool for in vitro toxicity screening of foods using biosensor-expressing human kidney cells

254th National Meeting (2017)

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