The development of whole smoke/aerosol exposure systems provides a means to conduct in vitro assessment of freshly generated whole smoke and aerosol from combustible and tobacco heating products (THP) as well as electronic nicotine delivery systems (ENDS). One challenge with such systems is ensuring sufficient throughput for in vitro toxicological studies in a timely manner. Vitrocell® has developed a high throughput whole smoke/aerosol exposure module designed to deliver concurrently up to seven different doses of smoke/aerosol and a clean air control to 48 wells of bacterial or mammalian cell cultures (six wells per dose). Characterization of this system was conducted with a series of experiments designed to assess smoke/aerosol delivery and biological responses from a Kentucky Reference 3R4F combustible cigarette or a commercially available THP. Dilution airflows consisting of 0.5 – 10 L/min for 3R4F and 0 (undiluted) - 4 L/min for the THP were evaluated. Smoke/aerosol deposition was quantified using fluorescence measurements (Ex 355/Em 485) of captured particulate matter and chemical analysis (e.g., glycerol, nicotine) of either DMSO (3R4F) or PBS (THP) traps within the module. Further characterization of the high throughput module was performed with the Neutral Red Uptake (NRU) assay to determine the cytotoxic response to 3R4F whole smoke. Current results demonstrate a dose-dependent deposition of smoke/aerosol constituents and a characteristic dose-dependent decrease in cell viability as indicated by the NRU assay.