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Updated: Jul 13, 2026
Technique for Isolation and Culture of Rat Jaw Bone Marrow Mesenchymal Stem Cells
Published on: May 31, 2024
[Osteogenic differentiation of rat bone marrow mesenchymal stem cell after transfection with recombinant pAd-bone
Meng-chun Qi1, Hong Sun, Jing Hu
1Department of Oral and Maxillofacial Surgery, West China School of Stomatology, Sichuan University, Chengdu 610041, China. drhu@vip.sohu.com
Objective:
To study the effect of recombinant pAd-BMP-7 on osteogenic differentiation of rat bone marrow mesenchymal stem cells (MSC).
Methods:
Recombinant pAd-bone morphogenetic protein (BMP) 7 was constructed and the titer of recombinant adenovirus was determined. pAd-BMP-7 and pAdTrack-CMV were used to transfect rat MSC. Transfection efficiency was measured by fluorescent microscope and BMP-7 expression was detected by RT-PCR and immunocytochemical analysis. The MSC were then randomly divided into 3 groups: group A received pAd-BMP-7 transfection, group B was transfected with pAdTrack-CMV, and group C received pAdTrack-CMV transfection plus bone supplements. Osteogenic differentiation of MSC was evaluated by examination of mineralization nodes formation.
Results:
The titer of pAd-BMP-7 reached about 2.0 x 10(15) pfu/L and transfection efficiency of exogenous gene was nearly 99% at day 2. The expression of exogenous gene sustained about 5 to 7 weeks, with a higher level during first 3 weeks. After transfection, transcription of BMP-7 and expression of BMP-7 protein were also verified in MSC. Compared with the negative results in group B, mineralization nodes were formed in both group A and group C. However, group A showed better formation of mineralization nodes than group C (P < 0.01).
Conclusions:
The results of this study indicated that recombinant pAd-BMP-7 can successfully transfect rat MSC and accelerate their osteogenic differentiation. The technique explored in this study provides a unique and valuable gene engineering approach for reconstruction of craniofacial bone defects.
