Background: The JUUL Nicotine Salt Pod System (NSPS) has no user modifiable settings and is temperature regulated to minimize the generation of combustion related byproducts. Aerosol generated from NSPS pods via a cotton wicking material was evaluated for cytotoxicity potential. Methods: Seven flavors (tobacco, mint/menthol, and fruit flavors; 18mg/mL nicotine) were evaluated. Cytotoxic potential was assessed using the NRU in vitro assay (OECD, TG 129; Labstat International ULC, Canada). CHO-WBL cells were treated with NSPS aerosol, positive control, or vehicle control for 24 hours utilizing a dimethyl sulfoxide (DMSO) extraction methodology (method T-502, Health Canada) with modifications. NSPS aerosol exposure occurred via separate dosing of aerosol collected mass in DMSO and gas vapor phase in CMF-PBS across a dose range of 0-1000 µg/mL. Additionally, cells were exposed to the sodium lauryl sulfate positive control at the concentration of 110 µg/mL. Cell viability following NSPS aerosol exposure was compared to the mainstream smoke component from a reference cigarette (3R4F) at dose range of 0-150 µg/mL. Results: No significant aerosol mediated toxicity was observed at any of the tested NSPS flavors or concentrations. EC50 for the NSPS aerosol and carrier control aerosol could not be calculated because cell viability was greater than 50% for all test conditions. Conclusions: Under the experimental conditions and based on the criteria for Evaluation of Cytotoxic Response (ISO 10993-5), all NSPS aerosol condensates generated from the 18mg/mL NSPS test articles were found to be non-cytotoxic.