Department of Orthopaedics, The First Affiliated Hospital of Bengbu Medical College, Laboratory of Tissue and Transplant in Anhui Province, Bengbu Medical College, Bengbu City, China.
Department of Orthopaedics, The Second Affiliated Hospital of Medical College, Shihezi University, Xinjiang, China.
Biomed Res Int. 2021 Jan 27;2021:7239783. doi: 10.1155/2021/7239783. eCollection 2021.
This study is aimed at investigating the effects of bone morphogenetic protein-7 (BMP-7) on the differentiation of bone marrow mesenchymal stem cells (BMSCs) into neuron-like cells . The rat BMSCs were isolated and identified, which were divided into the control, empty, recombinant rhBMP-7 transfection, and Lv-BMP-7 transfection groups. BMSCs were induced under different conditions. CCK-8 assay was performed to detect cell proliferation. ALP was used to detect cell activity. Cellular morphology after induction was observed. Immunofluorescence was conducted to detect the expression and location of nerve cell markers. Quantitative real-time PCR and Western blot analysis were performed to detect the mRNA and protein expression levels, respectively. The rhBMP-7 and Lv-BMP-7 promoted the proliferation of BMSCs, accompanied with increased ALP activities. Morphological observations revealed that rhBMP-7 and Lv-BMP-7 induced BMSCs to differentiate into neuron-like cells. Immunofluorescence revealed that the rhBMP-7 and Lv-BMP-7 groups showed positive expression of MAP-2 and Nfh in BMSCs. MAP-2 was mainly distributed in the cell body and cellular protrusion, while Nfh was mainly distributed in the cytoplasm and cell protrusion. Positive mRNA and protein expressions of MAP-2 and Nfh were observed in the cells of the rhBMP-7 and Lv-BMP-7 groups, and the expression levels were significantly higher than the control and empty groups. Both exogenous BMP-7 (rhBMP-7) and endogenous BMP-7 (Lv-BMP-7) can induce BMSCs to differentiate into neuron-like cells highly expressing the neuronal markers MAP-2 and Nfh.
本研究旨在探讨骨形态发生蛋白 7(BMP-7)对骨髓间充质干细胞(BMSCs)向神经元样细胞分化的影响。分离并鉴定大鼠 BMSCs,分为对照组、空载体组、重组 rhBMP-7 转染组和 Lv-BMP-7 转染组。在不同条件下诱导 BMSCs,采用 CCK-8 法检测细胞增殖,ALP 检测细胞活性,观察诱导后细胞形态,免疫荧光法检测神经细胞标志物的表达和定位,实时定量 PCR 和 Western blot 分析分别检测 mRNA 和蛋白表达水平。rhBMP-7 和 Lv-BMP-7 促进 BMSCs 增殖,同时 ALP 活性增加。形态学观察显示 rhBMP-7 和 Lv-BMP-7 诱导 BMSCs 分化为神经元样细胞。免疫荧光显示 rhBMP-7 和 Lv-BMP-7 组 BMSCs 中 MAP-2 和 Nfh 呈阳性表达。MAP-2 主要分布于细胞体和细胞突起,Nfh 主要分布于细胞质和细胞突起。rhBMP-7 和 Lv-BMP-7 组细胞中 MAP-2 和 Nfh 的阳性 mRNA 和蛋白表达均上调,表达水平明显高于对照组和空载体组。外源性 BMP-7(rhBMP-7)和内源性 BMP-7(Lv-BMP-7)均可诱导 BMSCs 分化为高表达神经元标志物 MAP-2 和 Nfh 的神经元样细胞。