OBJECTIVE To obtain ROS17/2.8 cell lines which were stably expressing EGFP reporter gene drived by COL1A1 promoter. METHOD A 3.6 Kb COL1A1 promoter from rat was cloned into pMD-18-T vector by PCR. This amplified promoter vector was digested to get several different length fragments which were then fused with EGFP reporter gene to construct eukaryotic expression vectors. ROS17/2.8 cell was stably transfected with these vectors by LipofectAMINE(TM) and selected by G418. RESULT The COL1A1-EGFP stably transfected cell lines were established. CONCLUSION The cell lines will be useful for studying the effects of microgravity on the activity of COL1A1 promoter and expression of gene related with bone form.