骨形态发生蛋白2抑制骨髓间充质干细胞成脂分化的优选浓度

发布时间：2018-07-18 17:30

【摘要】：背景:骨形态发生蛋白2既可以促进成骨,在不同浓度范围又可以调控骨髓间充质干细胞成脂分化的趋势。因此,骨形态发生蛋白2诱导骨髓间充质干细胞后,如果能够发现抑制其成脂的浓度范围,对于防治骨质疏松症具有重要意义。目的:确定骨形态发生蛋白2抑制骨髓间充质干细胞成脂分化的优选浓度。方法:(1)抽取大鼠股骨及胫骨的骨髓,体外用贴壁培养法获得原代骨髓间充质干细胞,流式细胞仪检测第2代细胞表面抗原标志物;(2)取第2及3代细胞,分为0.5,5,50,100,200μg/L骨形态发生蛋白2组及对照组。0.5,5,50,100,200μg/L骨形态发生蛋白2组骨髓间充质干细胞用成脂诱导液诱导3 d后,加入0.5,5,50,100,200μg/L骨形态发生蛋白2继续诱导14 d;对照组骨髓间充质干细胞成脂分化3 d,使用骨髓间充质干细胞培养基培养,未加骨形态发生蛋白2处理。结果与结论:(1)流式细胞仪鉴定培养的细胞为骨髓间充质干细胞;(2)油红O染色显示,5-50μg/L骨形态发生蛋白2诱导后,随骨形态发生蛋白2质量浓度的增加,脂滴数量增加;而100-200μg/L骨形态发生蛋白2诱导17 d后,随骨形态发生蛋白2质量浓度的增加,脂滴数量减少。说明100μg/L骨形态发生蛋白2可以抑制骨髓间充质干细胞成脂分化,从而减少大鼠脂肪细胞的形成,可以作为抑制骨髓间充质干细胞成脂分化的优选浓度。
[Abstract]:Background: bone morphogenetic protein 2 can promote osteogenesis and regulate the tendency of bone marrow mesenchymal stem cell adipogenesis in different concentration range. Therefore, bone morphogenetic protein-2 induced bone marrow mesenchymal stem cells, if the inhibition of its lipid concentration range can be found for the prevention and treatment of osteoporosis has important significance. Aim: to determine the optimal concentration of bone morphogenetic protein 2 (BMP 2) in inhibiting adipogenic differentiation of bone marrow mesenchymal stem cells (BMSCs). Methods: (1) the bone marrow of rat femur and tibia were extracted, primary bone marrow mesenchymal stem cells were obtained by adherent culture in vitro, the surface antigen markers of the second generation cells were detected by flow cytometry, and the second and third passage cells were obtained. The bone marrow mesenchymal stem cells (BMSCs) were induced with adipogenic fluid for 3 days. Bone morphogenetic protein-2 (BMP 2) was added to 0.5 渭 g / L for 14 d, and bone marrow mesenchymal stem cells (BMSCs) were cultured on bone marrow mesenchymal stem cells medium for 3 days without bone morphogenetic protein 2 (BMP 2) treatment. Results and conclusion: (1) the cultured cells were identified as bone marrow mesenchymal stem cells by flow cytometry (FCM), (2) oil red O staining showed that the number of lipid droplets increased with the increase of mass concentration of bone morphogenetic protein 2 (BMP 2) after induction of bone morphogenetic protein 2 (BMP 2). However, after induction of 100-200 渭 g / L bone morphogenetic protein 2 for 17 days, the number of lipid droplets decreased with the increase of mass concentration of bone morphogenetic protein 2. The results showed that 100 渭 g / L BMP 2 could inhibit the adipogenic differentiation of bone marrow mesenchymal stem cells, thus reducing the adipocyte formation in rats, and could be used as the optimal concentration for inhibiting the adipogenic differentiation of bone marrow mesenchymal stem cells.
【作者单位】：南昌大学第一附属医院骨科;
【基金】：江西省卫生厅科技计划课题(20141044)~~
【分类号】：R580

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