Refinement of a new method to estimate human bronchial epithelial cells ciliary beat frequency: effect of salmeterol

<bold>Introduction:</bold> In a previous work in bronchial cells from a Healthy donor and a COPD patient, we introduced a new method for the estimation of ciliary beat frequency (CBF). <bold>Aims of the study:</bold> To refine our previous method and to compare CBF values at 27-32-37°C (T) in Healthy cells vs Asthma cells in basal condition as well as following incubation with 1 µM salmeterol. <bold>Methods:</bold> Human bronchial epithelial cells from 1 healthy donor (H) and 1 patient suffering of asthma (A) were seed on inserts (“EpiAirways” from MatTek Corp.) which were put into a thermostated chamber at 37°-32° and 27°C and 5% CO2. Videos were recorded at 250 images/s during 2 s Salmeterol (1µM) was applied for 30 min, then cilia beating recorded again at different T. After that, cells were fixed on 4% paraformaldehyde then CBF recorded at 27°C. <bold>Results:</bold> The median CBF values were significantly lower in A cells vs H cells (p= 0.0001) at 27°C but not at 32° or 37°C (see Figure below). This difference at 27°C disappeared following treatment with salmeterol. When comparing the effects of salmeterol in H cells, we noticed an increase of CBF (p=0.08) at 37°C. <bold>Conclusions:</bold> We refined our method for CBF in human bronchial epithelial cells. At 27°C only, CBF in cells from A were lower than values in cells from H. Our results support the presence of T dependency of CBF values in ciliated cells from asthmatics. <fig><object-id>erj;64/suppl_68/PA2663/F1</object-id><object-id>F1</object-id><object-id>F1</object-id><graphic></graphic></fig>