Objective To explore an easier method to get epithelial cell and fibroblast from a rabbit' s trachea for trachea tissue engineering in vitro. Methods Digest the epithelial cell and fibroblast by trypsin after co-culture for 6 days. Distinguish these two kinds of cells according to their different tolerances to trypsin and culture characteristics. The new method and the traditional method were compared by the proliferation of the cells with cell counting and MIT assay. Results The purity of the cells got with this new method is up to 100% . The cells proliferation is as same as that cultured by traditional methods. Conclusion With this method, one can get two kinds of different cells from the original in one time, it is easier to practice than the traditional.