Objective:To establish a mouse preosteoblast cell lines overexpressing 11β-HSD1 gene. Methods:A lentiviral vector overexpressing mouse 11β-HSD1 was constructed. Lentivirus particles were generated in 293FT cells,followed by concentration and purification. MC3T3-E1 preosteoblast cells were infected by the lentivirus,which were later selected by BSD. The mRNA and protein expression of 11β-HSD1 was analyzed by real-time PCR and Western blot,respectively. MC3T3-E1 osteogenic differentiation was detected using Alizarin red staining. Results:The lentiviral vector of mouse 11β-HSD1 overexpression was successfully constructed,and the MC3T3-E1 preosteoblast cell lines were infected efficiently by the lentivirus. The mRNA and protein levels of 11β-HSD1 were higher in MC3T3-E1 cells infected by 11β-HSD1 overexpression lentivirus than that of cells infected by the negative control lentivirus. Alizarin red staining showed that the differentiation ability toward osteoblasts of MC3T3-E1 cells was not influenced by lentivirus infection. Conclusion:Our findings showed that 11β-HSD1 gene was effectively overexpressed in MC3T3-E1 preosteoblast cells using lentivirus infection,which provides a useful tool for further studies of 11β-HSD1 function in osteoblasts.