We attempted to determine the growth characteristics of cultured lung fibroblasts of layer type chickens and to investigate presence of matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) in these cells in vitro. Lung fibroblasts were isolated, characterized and subcultured from one-day-old layer type chicken lungs. Two different methods, explant culture and enzymatic techniques, were used for culturing and the results were compared. The presence of MMP-2 and TIMP-1 was shown in cultured fibroblasts by immunocytochemical staining, immune blotting and zymography methods. Immune expressions of neither MMP-9 nor TIMP-2 enzymes could be detected.