Cannabis is commonly consumed via cannabis smoke which contains combustion products. There is an increasing interest in vaporizers, which heat the cannabis flower without burning. Cannabis smoke extracts (CaSE) for in vitro tests are traditionally made with media containing 30% methanol. We optimize methods for cannabis smoke and vapour extracts (CaVE) for submerged cell cultures and whole cannabis exposures for air-liquid interface (ALI) cultures.

CaSE was produced traditionally, in ethanol, or in methanol. CaVE was created by bubbling cannabis vapor through ethanol. Combustion products captured were assessed by optical density at 320nm. Cannabinoids were assessed using high-performance liquid chromatography. Cytotoxicity of extracts was assessed in A549 cells by LDH release. ALI cultures were exposed to cannabis vapor using the SCIREQ expoCube.

Traditional CaSE contained 6.77ug/ml d9-THC; meanwhile, CaSE in ethanol contained 436.7ug/ml d9-THC and methanol contained 302.8ug/ml d9-THC. CaSE prepared in ethanol and methanol captured 9 and 7 more minor cannabinoids than traditional CaSE. Ethanol and methanol captured 5.1x and 3.8x the combustion products of traditional CaSE. CaVE produced in ethanol captured 83.8 ug/ml of d9-THC and contained 3 minor cannabinoids. CaSE with ethanol caused 50% cytotoxicity at 200ng/ml d9-THC while CaVE caused 50% cytotoxicity at 1000ng/ml of d9-THC. ALI cultures exposed to cannabis vapor show higher levels of cytotoxicity compared to submerged cultures.

We show a reliable method to capture a higher quantity and more diverse cannabinoid profile for in vitro experimentation. Overall, this work will set the foundation for future in vitro work in inhaled cannabis products.