Preparation and Culture of Precision-Cut Lung Slices

Healthy human lung tissue was obtained from Donor Alliance, and IPF tissue was obtained from lung biopsies or explanted lungs from confirmed cases of IPF (IRB 11-1664) as described previously (74 (link)). Tissue from patients with silicosis was obtained after lung transplant as described previously (75 (link)). For the generation of PCLS, freshly explanted healthy or IPF lungs were kept on ice and processed within 24 hours. Peripheral sections of lungs were inflated with low–melting point agarose (MilliporeSigma) by cannulation of a visible bronchus; then 6 × 6 × 6 mm cubes were cut from the lung, embedded in low–melting point agarose, and sectioned using a vibratome (Leica). We cultured 300 μm slices in Eagle minimal essential medium (EMEM, Lonza) with 10% heat-inactivated fetal bovine serum (FBS, Atlanta Biologicals) and 1% penicillin/streptomycin/l-glutamine with or without ABT-263 (5 μM, Medchemexpress) for 24 hours.