Citation:

Winters, B., M. Angrish, A. Wallace, J. Pleil, AND M. Madden. In Vitro Toxicity Screening Technique for Volatile Substances Using Flow-Through System##. Gordon Research Conference-Advanced Investigation in Mechanistic Toxicology, Andover, NH, August 13 - 18, 2017.

Impact/Purpose:

This presentation is to an audience of cellular toxicologists describing a new xenobiotic exposure system which will allow toxicity testing in a quick and inexpensive manner. The responses of the cells in testing system are being validated with chemicals that have been studied extensively in humans in vivo. The presentation of this proposed system of testing will provide the audience with a chance to comment on the rationale of the goals and the design of the system.

Description:

In 2007 the National Research Council envisioned the need for inexpensive, rapid, cell based toxicity testing methods relevant to human health. Recent advances in robotics, automation, and miniaturization have been used to address this challenge. However, one drawback to currently-available approaches is that many of these chemicals are volatile and not amenable to liquid-based in vitro assays. We assembled an in vitro cell culture apparatus capable of screening volatile chemicals for toxicity with potential for miniaturization for higher throughput using a metabolomics approach. BEAS-2B lung cells were grown in an enclosed culture apparatus under air-liquid interface (ALI) conditions, for exposure to both gaseous and insoluble chemicals. Additionally, use of ALI conditions allows for release of endogenously-produced gaseous molecules without interference by medium on the apical surface. To identify potential xenobiotic-induced perturbations in cell homeostasis, we monitored for alterations of endogenously-produced gaseous molecules in air directly above the cells, termed “headspace”. Alterations in specific endogenously-produced gaseous molecules (e.g., signaling molecules nitric oxide (NO) and carbon monoxide (CO)) in headspace is indicative of xenobiotic-induced perturbations of specific cellular processes. Additionally, endogenously-produced volatile organic compounds (VOCs) may be monitored in a nonspecific, discovery manner to determine whether cell processes are perturbed by xenobiotic exposure, potentially producing specific patterns of changes. Preliminary results indicate the novel cell culture system is capable of detecting gaseous products of specific enzyme pathways such as CYP2A6. Numerous VOCs were detected in the headspace of cells grown in the novel cell culture apparatus. The system is being optimized to minimize artifact volatiles using different materials. We believe our novel cell culture apparatus, once refined and validated, will allow for screening of both volatile and non-volatile xenobiotics by measuring cellular responses detected as alterations in gaseous molecules released by cells. [This abstract may not reflect official US EPA policy.]

Record Details: