Knockoutof nipbl gene down-regulates the abilities of proliferation and osteogenic differentiation in mouse bone marrow mesenchymal stem cells / 中国组织工程研究

ABSTRACT

BACKGROUND: Cornelia de Lange syndrome is a genetic disease with multiple developmental defects, of which NIPBL is the main pathogenic gene. OBJECTIVE: To investigate the effect of NIPBL gene on the proliferation and osteogenic differentiation of bene marrow mesenchymal stem cells. METHODS: The NIPBL+/- mice were constructed by NIPBL-Loxp and Cre mice and used as experimental group, and the wild-type NIPBL+/+ mice served as control group. Mouse bone marrow mesenchymal stem cells were isolated and cultured in the two groups. Cell proliferation was detected using cell counting kit-8 assay when thecells were passed to the third generation. Osteoblastic differentiation was then compared between two groups after osteogenesis induction. RESULTS AND CONCLUSION: The proliferation capacity of bone marrow mesenchymal stem cells in the experimental group was lower than that in the control group (P < 0.05). The activity of alkaline phosphatase in the experimental group was significantly lower than that in the control group on the 7th day of osteogenic induction (P < 0.05). The expression levels of osteogenic genes and proteins (Runx2 and OCN) in the experimental group were lower than those in the control group after osteogenic induction (P < 0.05). On the 21st day of osteogenic induction, results from alizarin red staining indicated there were more red calcium nodules in the control group than the experimental group under inverted microscope. These findings suggest that NIPBL gene knockout can reduce the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells.