A novel apparatus for the exposure of cultured cells to volatile agents.

摘要

This article presents a novel exposure apparatus that allows the exposure of cultured cells to volatile chemicals, e.g., inhalation anesthetics. The apparatus consists of an exposure chamber and a tightly linked vaporizer unit with pumps and valves allowing adjustable fluxes of mixtures of test chemicals and carrier gas under open and closed-circuit conditions. The exposure chamber uses commercially available cell culture flasks and accommodates up to 12 flasks simultaneously. Both modules fit into a standard culture incubator. The exposure chamber may be mounted onto an oscillating axis to tilt the cultures periodically forth and back, thus allowing direct contact of the cells with test atmosphere. The vaporizer unit is connected to a personal computer which lets the experimenter set the "open" and "close" intervals of individual valves thereby controlling the composition and flow rate of the test gas mixture. The vapor concentration of test chemicals can be monitored at the inlet and outlet using infrared photodetectors or mass spectrometers. Computer-aided processing of exposure protocols allows unattended runs. Exposure protocols can be scripted and stored on disk, thus ensuring interexperimental reproducibility of complex exposure profiles. As an application example, the effect of three volatile anesthetics, halothane, enflurane, and isoflurane, on the viability of three commercially available cell lines (A549--human lung carcinoma, HTC-rat hepatoma, MDCK--Madin-Darby canine kidney) was investigated. After exposure to haloalkyl vapors (3%) for 6 and 24 h, respectively, significantly increased LDH levels versus controls, indicating cellular membrane damage, were detected in A549 and hepatoma cells after exposure for 24 h. Hepatoma cells showed a significant LDH release also after 6 h exposure to isoflurane. On the other hand, LDH release from MDCK cells was not significantly different from controls even after 24 h of continuous exposure to any of the tested anesthetics.