Objective To establish T739 mouse bladder cancer cell lines and tumor-bear model stably expressing human MUC1. Methods The cell lines and animal model were established by Lipofectamine-mediated transfection of human MUC1 full length cDNA into BST739 cells, followed by C,418 selection and colonel culture. PCR, RT-PCR, FCM and immunohistochemistry were carried out to confirm the stable integration of MUC1 cDNA into the mouse genomo and expression of MUC1 mRNA and proteins in the cells. The growth characteristics of BST739-MUC1 were also investigated in vitro and in vivo. Results The MUC1 was stably integrated into the cell genome and expressed in cell membrane and plasma. The growth characteristics of transfected cells did not alter in vitro and in vivo. Conclusion T739 mouse bladder cancer cell lines and tumor-bear model expressing human MUC1 were successfully established for the development of tumor vaccines targeting MUC1.