Knockout of NIPBL gene down-regulates the abilities of
proliferation and osteogenic differentiation in mouse bone marrow mesenchymal
stem cells

doi:10.3969/j.issn.2095-4344.2029

Abstract

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 the cells 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 7^th 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 21^st 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.

Key words: NIPBL gene, developmental defect, bone marrow mesenchymal stem cells, cell proliferation, cell differentiation, osteogenic induction, calcium nodules, the National Natural Science Foundation of China

CLC Number:

Lin Ming, Pan Jinyong, Zhang Huirong. Knockout of NIPBL gene down-regulates the abilities of proliferation and osteogenic differentiation in mouse bone marrow mesenchymal stem cells[J]. Chinese Journal of Tissue Engineering Research, 2020, 24(7): 1002-1008.

Figures/Tables 9

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