Objective:To establish fibroblasts stably expressing human cementum protein 1 (hrCEMP1). Methods: Eukaryotic expression vector pcDNA3.1-hrCEMP1 was transduced into NIH3T3 cells using SofastTM,and stably transfecting cell line was selected by G418. The transcription of hrCEMP1 in the transfecting cells was determined by RT-PCR and enzyme-linked immunosorbent assay(ELISA). Results: hrCEMP1 mRNA was transcripted and expressed in the transfected NIH3T3 cells. Conclusion:With positive compound transfection, outside hrCEMP1 gene can be successfully transducted and expressed in NIH3T3 cells, which is the foundation of researching the induction of hrCEMP-1 on fibroblast cells.