Human lung fibroblast isolation

Lung fibroblasts were derived from surgical lung biopsies (SLBs) from patients who exhibited slowly or rapidly progressing IPF (slow IPF and rapid IPF, respectively) over the first year after diagnosis¹². Primary normal lung fibroblasts were derived from non-fibrotic lung samples lacking any evidence of disease. Histologically-proven IPF and normal lung samples were dissociated mechanically as previously described in detail¹³. Briefly, SLBs were mechanically dispersed in 150 cm³ flasks (Corning), combined with Dulbecco’s modified Eagle Medium (Lonza), containing 15% fetal bovine serum (Cell Generation), 0.1% Pen/Strep/Amphotericin B (10,000 U of penicillin/ml, 10,000 μg of streptomycin/ml, and 25 μg Amphotericin B/ml) (Lonza), 0.1% (29.20 mg/mL) L-glutamine (Corning), and Primocin at 100 μg/ml (Invivogen). Isolated fibroblasts were incubated at 37 °C in 10% CO₂ until confluency was reached. The cells were passaged 3 to 4 times before use in the experiments described herein. In the present study, we analyzed a total of 4 normal fibroblast lines, 7 fibroblast lines derived from IPF patients who showed slow progression, and 7 fibroblast lines derived from IPF patients who showed rapid progression.