Platelet-derived growth factor-BB induces the differentiation of rat bone marrow mesenchymal stem cells into osteoblasts

doi:10.3969/j.issn.2095-4344.2200

Abstract

Abstract: BACKGROUND: It has been shown that platelet-derived growth factor-BB secreted by mouse osteoclasts can promote the formation of osteoblasts and vascular cells. Whether platelet-derived growth factor-BB can promote the differentiation of bone marrow mesenchymal stem cells into osteoblasts has not been studied.
OBJECTIVE: To investigate the feasibility of platelet-derived growth factor-BB on inducing bone marrow mesenchymal stem cells of rats into osteoblasts.
METHODS: (1) The bone marrow mesenchymal stem cells of Sprague-Dawley rats were isolated and cultured in vitro. The surface markers CD45, CD29 and CD34 of the third generation were identified by flow cytometry. Bone marrow mesenchymal stem cells with high purity were selected for platelet-derived growth factor-BB intervention experiments. (2) Bone marrow mesenchymal stem cells at passage 3 were induced by 25 μg/L platelet-derived growth factor-BB for 3 days. Cellular immunofluorescence was used to identify type I collagen, followed by alkaline phosphatase modified calcium-cobalt method and azo coupling method kit, alizarin red method. The surface characteristics of osteoblasts were identified. Finally, RT-PCR was used to detect the mRNA levels of Runx-2 and Gli-2 genes.
RESULTS AND CONCLUSION: (1) Cellular immunofluorescence staining showed that the green fluorescence expression rate of type I collagen was more than 90% after induction. (2) Alkaline phosphatase modified calcium-cobalt method demonstrated that black cords were attached to the cytoplasm of the cells, indicating that these cells expressed active alkaline phosphatase. Alkaline phosphatase azo coupling method showed that blue active alkaline phosphatase was found in the cytoplasm of cells. Alizarin red staining exhibited that induced cells gathered together to form small red calcium nodules. (3) RT-PCR showed that the mRNA levels of Gli-2 and Runx-2 genes were higher than those in bone marrow mesenchymal stem cells after platelet-derived growth factor-BB induction (P < 0.05). (4) It is concluded that rat bone marrow mesenchymal stem cells can be differentiated into functional osteoblasts after being induced by platelet-derived growth factor-BB.

Key words: stem cells, bone marrow mesenchymal stem cells, platelet-derived growth factor BB, osteoblasts, calcium nodules, alkaline phosphatase

CLC Number:

Wei Qin, Zhang Xue, Ma Lei, Li Zhiqiang, Shou Xi, Duan Mingjun, Wu Shuo, Jia Qiyu, Ma Chuang. Platelet-derived growth factor-BB induces the differentiation of rat bone marrow mesenchymal stem cells into osteoblasts[J]. Chinese Journal of Tissue Engineering Research, 2021, 25(19): 2953-2957.

Figures/Tables 5

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