Precision cut lung slices: a novel versatile tool to examine host–pathogen interaction in the chicken lung

摘要

PCLS viability was assessed by AlamarBlue assay ( and ) and Live/Dead staining ( ). PCLS (500 μm) prepared from -eGFP transgenic chickens were cultured for 7 days in either DMEM/F12/FCS (blue circle), DMEM/F12 (red triangle) or DMEM (black square) and viability was assessed using the AlamarBlue assay ( ).  = 6–18 slices generated from 3 birds, data are represented as the mean ± SD. PCLS prepared from SPF PA12 chickens (200–300 μm) were cultured for 7 days in either DMEM/F12/FCS (blue circle), DMEM/F12 (red triangle) or DMEM (black square) and were stained with a Live/Dead kit and subsequently examined by epifluorescence microscopy ( ). The number of dead cells were enumerated in images from 3 to 4 PCLS generated from 2 birds at different time-points using the Fiji-ImageJ image processing software. Data are expressed as the mean ± SD, per time-point of one representative experiment.  = 3–4 slices. The maintenance of cell viability in PCLS (500 μm) prepared from -eGFP transgenic chickens was assessed in DMEM media by AlamarBlue assay ( ).  = 16 slices generated from 2 birds, data are represented as the mean ± SD.