D. Ritter et al., In vitro exposure of isolated cells to native gaseous compounds - Development and validation of an optimized system for human lung cells, EXP TOX PAT, 53(5), 2001, pp. 373-386
An exposure system for adherent growing cells to native gaseous compounds w
as developed using air/liquid culture techniques on the basis of the Cultex
system(1). In contrast to other exposure systems the reproducible testing
of native environmentally relevant gases without changing their physical or
chemical properties including heating, CO2-content and humidity is possibl
e. Specially designed systems for medium flow and gas support guarantee the
nutrification and humidification as well as the direct gas contact of the
exposed cells which are cultivated on microporous membranes (0.4 mum pore s
ize), The system works independently of a cell culture incubator offering t
he possibility to analyze any relevant gas mixture directly under indoor or
outdoor conditions.
Several experimental approaches were carried out to characterize the proper
ties of the system. In exploratory experiments without cells, the reproduci
bility and quality of the gas/membrane contact could be demonstrated. Expos
ures of human lung fibroblasts (Lk004 cells) and human lung epithelial cell
s (HFBE-21 cells) to synthetic air, ozone (202 ppb, 510 ppb) and nitrogen d
ioxide (75 ppb to 1200 ppb) established that cells could be treated for 120
minutes without significant loss of cellular viability. At the same time,
the experiments confirmed that such exposure times are long enough to detec
t biological effects of environmentally relevant gas mixtures. The analysis
of viability (viable cell number, tetrazoliumsalt cleavage) and intracellu
lar end-points (oxidized/reduced glutathione, ATP/ADP) showed that both gas
es induced relevant cellular changes. In summary, the efficiency and practi
cability of this newly developed exposure system for adherent human lung ce
lls could be clearly demonstrated.