Twenty five heart-lung blocs were isolated from adult male Lewis rats weighting 180±15g. Organs were mounted in bioreactor and treated with modified detergent-enzymatic decellularization protocol with tracheal ventilation. Decellularization of native lungs was histologically characterized. Specific matrix components were analyzed by immunohistochemistry staining. Ultrastructural analysis of lung matrix was performed by scanning electronic microscopy. DNA content in native and decellularized lungs was evaluated by spectrophotometer. Recellularization of lung scaffold was carried out by perfusion with oxygenated medium with mesenchymal stromal cells. A cell proliferation assay and life/dead staining were used to measure viability and metabolic activity. Acellular lungs maintained native ultrastructual proprieties with extracellular matrix proteins preservation. About 92% of DNA was removed after treating.Results of cell proliferation assays showed cells viability and metabolic activity during cultivation on the scaffold.Modified lung decellularization protocol allows cell attachment for recellularized rat lung obtainment.