Primary fibroblast cell cultures from transbronchial biopsies of lung transplant recipients

Background Survival after lung transplantation is limited by the developmen t of bronchiolitis obliterans (BO) that is a fibroproliferative process and regarded as the histological marker of chronic rejection, To study further the pathogenesis of BO we attempted to establish primary fibroblast cell c ultures from transbronchial lung biopsies (TBBs)of lung transplant recipien ts. Methods. One to two TBB samples from each patient were collected in sterile phosphate-buffered saline. Biopsies were cut into small pieces and placed onto 25-cm(2) culture flasks for cell culture and kept under standard cell culture conditions (21% O-2, 5% CO2, 37 degreesC). Culture medium consisted of RPMI 1640, 10% fetal calf serum, L-glutamine, HEPES, and antibiotics. A fter reaching confluence, fibroblasts were passaged into 75-cm(2) flasks. Results. The success rate of establishing fibroblast cultures from transbro nchial lung biopsies was 54% (27/50). Cell growth was independent of patien t age, transplant type, underlying lung disease, indication for transbronch ial lung biopsies, grade, or type of rejection and infection. Conclusions. We have established a novel method of culturing fibroblasts fr om lung transplant recipients. We consider this method as an unique human i n vitro model to study the pathogenetic mechanisms leading to BO.