A new approach to evaluate toxicity of gases on mobile cells in culture.
A novel technique is described that measures the degree of toxicity of short-term exposure to gaseous pollutants or other chemical compounds on cultured cells, in 30 min. This technique, based on the study of the mobility properties of activated macrophages, consists of an image analysis procedure incorporating a specific exposure chamber (EC). The EC, which is developed from commercial culture flasks (50 ml, 25 cm(2) of culture surface), was first used to maintain cells in culture conditions, overnight, prior to the assay. In order to measure toxicity, it was then connected to the gaseous pollutant or chemical source. After exposing the culture medium and cells to the gas stream for 10 min, fMLP, a chemotactic factor, was added and the mobility of the macrophages measured by superimposing sequential analogue images captured by a CCD camera that were digitised and analysed using a software developed for this purpose. For example, the effect of ozone on macrophage-like cell (THP-1) was investigated. After exposure to 0.1 and 0.5 ppm, cells lost, respectively 79% and 90% of their mobility, compared to the control sample.
Laval-Gilly
P
P
Département Génie Biologique-Génie de l'Environnement, Institute de Technologies de Metz, Impasse Alfred Kastler, 57970, Yutz, France. plg@iut.univ-metz.fr
Falla
J
J
Klestadt
D
D
Henryon
M
M
D001273:Atmosphere Exposure Chambers; D002465:Cell Movement / Q000187:drug effects* / Q000502:physiology; D002478:Cells, Cultured; D005740:Gases / Q000633:toxicity*; D006801:Humans; D007091:Image Processing, Computer-Assisted; D008264:Macrophages / Q000166:cytology / Q000187:drug effects* / Q000502:physiology; D010126:Ozone / Q000633:toxicity; D013997:Time Factors
D005740:Gases; D010126:Ozone