盐酸四环素对大鼠骨髓间充质干细胞的成骨诱导分化活性
发布时间：2018-11-28 17:09
【摘要】：背景:盐酸四环素不仅具有促进骨髓间充质干细胞增殖的作用,还具有极强的骨亲活性。研究盐酸四环素诱导骨髓间充质干细胞向成骨细胞分化,可为获得骨组织工程细胞提供新方法。目的:观察体外盐酸四环素对大鼠骨髓间充质干细胞向成骨细胞分化的作用。方法:骨髓全血细胞贴壁法分离培养大鼠骨髓间充质干细胞,通过细胞形态和骨髓间充质干细胞标志物CD34、CD45、CD90、CD29流式检测对骨髓间充质干细胞进行鉴定;取第3代骨髓间充质干细胞与盐酸四环素共培养,以含体积分数10%FBS的L-DMEM的完全培养基培养做对照组。通过检测碱性磷酸酶活性、Ⅰ型胶原和骨钙素免疫组织化学染色、茜素红矿化钙结节组织化学染色、Ⅰ型胶原蛋白和骨钙素的RT-PCR的基因表达,研究盐酸四环素对骨髓间充质干细胞的成骨诱导分化活性。结果与结论:①盐酸四环素能够持续促进骨髓间充质干细胞的碱性磷酸酶活性增加,与对照组比较具有显著性差异;②盐酸四环素诱导培养骨髓间充质干细胞21 d,Ⅰ型胶原蛋白和骨钙素染色表达阳性,茜素红染色出现红色钙结节,RT-PCR检测Ⅰ型胶原蛋白和骨钙素基因表达;对照组程阴性表达;③结果表明,盐酸四环素具有明显促进骨髓间充质干细胞成骨诱导分化的作用。
[Abstract]:BACKGROUND: Tetracycline hydrochloride not only has the effect of promoting the proliferation of mesenchymal stem cells in the bone marrow, but also has extremely strong bone affinity. To study the differentiation of bone marrow mesenchymal stem cells into osteoblasts by tetracycline hydrochloride, a new method can be provided for obtaining bone tissue engineering cells. Objective: To observe the effect of tetracycline hydrochloride on the differentiation of bone marrow mesenchymal stem cells into osteoblasts. Methods: The bone marrow mesenchymal stem cells were isolated and cultured by bone marrow whole blood cell wall-wall method, and the bone marrow mesenchymal stem cells were identified by the cell morphology and the bone marrow mesenchymal stem cell markers CD34, CD45, CD90 and CD29. The 3-generation bone marrow mesenchymal stem cells were co-cultured with tetracycline hydrochloride, and the control group was cultured with a total medium of L-DMEM with a volume fraction of 10% FBS. By detecting the activity of alkaline phosphatase, the immunohistochemical staining of type I collagen and bone calcium, the histochemical staining of I-type collagen and bone calcium, the expression of the gene of the RT-PCR of type I collagen and bone calcium, the bone-induced differentiation activity of tetracycline on the mesenchymal stem cells of the bone marrow was studied. Results and Conclusion: Tetracycline hydrochloride can continuously promote the increase of the alkaline phosphatase activity of the mesenchymal stem cells in the bone marrow, and there is a significant difference between the control group and the control group, and the positive expression of the bone marrow mesenchymal stem cells 21d, the type I collagen and the bone calcium in the tetracycline-induced culture of the tetracycline hydrochloride is positive, The red calcium nodules and RT-PCR were used to detect the expression of the type 鈪，
本文编号：2363639
[Abstract]:BACKGROUND: Tetracycline hydrochloride not only has the effect of promoting the proliferation of mesenchymal stem cells in the bone marrow, but also has extremely strong bone affinity. To study the differentiation of bone marrow mesenchymal stem cells into osteoblasts by tetracycline hydrochloride, a new method can be provided for obtaining bone tissue engineering cells. Objective: To observe the effect of tetracycline hydrochloride on the differentiation of bone marrow mesenchymal stem cells into osteoblasts. Methods: The bone marrow mesenchymal stem cells were isolated and cultured by bone marrow whole blood cell wall-wall method, and the bone marrow mesenchymal stem cells were identified by the cell morphology and the bone marrow mesenchymal stem cell markers CD34, CD45, CD90 and CD29. The 3-generation bone marrow mesenchymal stem cells were co-cultured with tetracycline hydrochloride, and the control group was cultured with a total medium of L-DMEM with a volume fraction of 10% FBS. By detecting the activity of alkaline phosphatase, the immunohistochemical staining of type I collagen and bone calcium, the histochemical staining of I-type collagen and bone calcium, the expression of the gene of the RT-PCR of type I collagen and bone calcium, the bone-induced differentiation activity of tetracycline on the mesenchymal stem cells of the bone marrow was studied. Results and Conclusion: Tetracycline hydrochloride can continuously promote the increase of the alkaline phosphatase activity of the mesenchymal stem cells in the bone marrow, and there is a significant difference between the control group and the control group, and the positive expression of the bone marrow mesenchymal stem cells 21d, the type I collagen and the bone calcium in the tetracycline-induced culture of the tetracycline hydrochloride is positive, The red calcium nodules and RT-PCR were used to detect the expression of the type 鈪，
本文编号：2363639
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