Paracrine effects of bone marrow mesenchymal stem cells on biological function of osteoblasts

doi:10.3969/j.issn.2095-4344.2014.10.001

Abstract

Abstract:

BACKGROUND: Some studies have shown that bone marrow mesenchymal stem cells have a certain role in the repair of bone defects, but the clinical application is limited because of ischemia, hypoxia and inflammation of injured tissues.
OBJECTIVE: To investigate the paracrine effects of bone marrow mesenchymal stem cells on osteoblast biological function.
METHODS: Bone marrow mesenchymal stem cells were isolated using standard Ficoll-Paquedensity gradient centrifugation. Mesenchymal stem cell conditioned medium was prepared to cultivate osteoblasts, MG63. Proliferation of MG63 cells was analyzed by cell counting kit-8. Migration of MG63 cells was analyzed by cell scratch method. Alkaline phosphatase activity of MG63 cells was analyzed by microplate test kit. Real-time PCR was performed to evaluate osteoblast differentiation markers, alkaline phosphatase, collagen type I and osteocalcin. Alizarin red staining was performed to evaluate osteoblast mineralization.
RESULTS AND CONCLUSION: The cells were strongly positive for CD44, CD73 and CD90, but negative for CD34. MG63 cells cultured in the conditioned medium showed better proliferation and migration than those cultured in the Dulbecco’s modified Eagle’s medium. The activity and mRNA expression of alkaline phosphatase were much higher after induction of 4, 7 days (P < 0.01). There was no significant difference in expression of collagen type I and osteocalcin after induction of 4 days, but they were significantly higher than those in the control group after induction of 7 days (P < 0.05). Alizarin red staining showed that the number of calcium nodules was increased and the mineral apposition was enhanced after induction of 21 days with the conditioned medium. These findings suggest that the paracrine substance of bone marrow mesenchymal stem cells can significantly promote osteoblast proliferation, migration, differentiation and mineralization.

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

全文链接：

Key words: bone marrow, mesenchymal stem cells, culture media, conditioned, osteoblasts, cell proliferation, cell differentiation, cell migration assays

CLC Number:

R394.2

Li Cheng, Zhou Hai-bin. Paracrine effects of bone marrow mesenchymal stem cells on biological function of osteoblasts [J]. Chinese Journal of Tissue Engineering Research, 2014, 18(10): 1477-1483.

Figures/Tables 4

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