Bone marrow mesenchymal stem cells transfected with recombinant adenovirus vectors carrying basic fibroblast growth factor in co-culture with ligament fibroblasts

doi:10.3969/j.issn.2095-4344.2015.28.001

Abstract

Abstract:

BACKGROUND: Basic fibroblast growth factor (bFGF) plays an important role in the ligament tissue healing process, and bone marrow mesenchymal stem cells transfected with growth factors can be used as seed cells in ligament tissue engineering.

OBJECTIVE: To observe biological effect of bone marrow mesenchymal stem cells transfected with recombinant adenovirus vectors carrying bFGF in three-dimensional co-culture with ligament fibroblasts.

METHODS: Passage 3 bone marrow mesenchymal stem cells were divided into three groups: control group, Ad-EGFP group and Ad-bFGF group. Under a phase contrast microscope, the changes in cell morphology were observed and the rate of transfection was analyzed by flow cytometry. Proliferation of bone marrow mesenchymal stem cells and ligament fibroblasts was analyzed by MTS, the expression of bFGF protein in bone marrow mesenchymal stem cells was determined by ELISA. Scleraxis, collagen type I, collagen type III, decorin and cartilage oligomeric matrix protein levels were detected in BMSCs and ligament fibroblasts using real-time fluorescent quantitative PCR.

RESULTS AND CONCLUSION: Recombinant adenovirus-mediated bFGF gene could transfect bone marrow mesenchymal stem cells efficiently. After co-culture for 3, 6 days, compared with the control group and Ad-EGFP group, in the Ad-bFGF group, the proliferation ability of bone marrow mesenchymal stem cells and ligament fibroblasts was enhanced (P < 0.01), the expression of bFGF protein in supernatant was obviously higher (P < 0.01), the collagen type I, collagen type III, decorin and cartilage oligomeric matrix protein mRNA expression decreased in the ligament fibroblasts (P < 0.01), but the mRNA expression of Scleraxis, collagen type I, collagen type III in bone marrow mesenchymal stem cells increased (P < 0.01). The results suggest that the co-culture of Ad-bFGF-transfected bone marrow mesenchymal stem cells with ligament fibroblasts promotes the proliferation of ligament fibroblasts while decreases the collagen synthesis at the same time, and stimulates the proliferation and differentiation of bone marrow mesenchymal stem cells into ligament fibrolasts.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Bone Marrow, Mesenchymal Stem Cells, Patellar Ligament, Fibroblasts

CLC Number:

R394.2

Li Bin, Chen Liao-bin, Qi Yong-jian, Hu Dong-cai, Chen Biao. Bone marrow mesenchymal stem cells transfected with recombinant adenovirus vectors carrying basic fibroblast growth factor in co-culture with ligament fibroblasts[J]. Chinese Journal of Tissue Engineering Research, 2015, 19(28): 4429-4434.

Figures/Tables 2

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