Idiopathic pulmonary fibrosis (IPF) is a fatal disease of the lower respiratory tract with
restricted therapeutic options. Repetitive injury of the bronchoalveolar epithelium leads
to activation of pulmonary fibroblasts, differentiation into myofibroblasts and excessive
extracellular matrix (ECM) deposition resulting in aberrant wound repair. However,
detailed molecular and cellular mechanisms underlying initiation and progression of
fibrotic changes are still elusive. Here, we report the generation of a representative
fibroblast reporter cell line (10-4ABFP) to study pathophysiological mechanisms of IPF in
high throughput or high resolution in vitro live cell assays. To this end, we immortalized
primary fibroblasts isolated from the distal lung of Sprague-Dawley rats. Molecular
and transcriptomic characterization identified clone 10-4A as a matrix fibroblast
subpopulation. Mechanical or chemical stimulation induced a reversible fibrotic state
comparable to effects observed in primary isolated fibroblasts. Finally, we generated a
reporter cell line (10-4ABFP) to express nuclear blue fluorescent protein (BFP) under
the promotor of the myofibroblast marker alpha smooth muscle actin (Acta2) using
CRISPR/Cas9 technology. We evaluated the suitability of 10-4ABFP as reporter tool in
plate reader assays. In summary, the 10-4ABFP cell line provides a novel tool to study
fibrotic processes in vitro to gain new insights into the cellular and molecular processes
involved in fibrosis formation and propagation
