Coding sequences of BMP-2 and BMP-7 were amplified using PCR and ligated with a DNA sequence encoding a flexible peptide (Gly4Ser)5. The fusion gene was inserted into plasmid pIRESneo3. The expression level of BMP2/7 heterodimers in the transfected CHO-K1 cells was 230.75+/-13.34 ng/mL. Culture medium of stably tansfected clone pool was collected as conditional medium to treat osteoblast MC3T3 cells. Staining of Alkalin phosphatase and Alizarin red demonstrated that the conditional medium significantly promoted osteogenic differentiation to a higher extent than BMP-2 homodimers expressed in either CHO-K1 cells or E. coli. Transcriptional levels of Osteogenic phenotype-related molecular markers such as OC, ALP, Runx2 and Osx were increased (P<0.05), and BMP/Smad signal activities were significantly enhanced by BMP-2/7 heterodimers, comparing with BMP-2 homodimers (P<0.05). The results demonstrate that BMP-2/7 heterodimers expressed in CHO-K1 cells have potent ability to stimulate osteogenic differentiation.
3T3 Cells ; Animals ; Artificial Gene Fusion ; Bone Morphogenetic Protein 2 ; biosynthesis ; genetics ; Bone Morphogenetic Protein 7 ; biosynthesis ; genetics ; CHO Cells ; Cell Differentiation ; drug effects ; Cloning, Molecular ; Cricetinae ; Cricetulus ; Dimerization ; Escherichia coli ; genetics ; metabolism ; Gene Fusion ; genetics ; Humans ; Mice ; Osteoblasts ; cytology ; Osteogenesis ; drug effects ; Transfection