首页> 外文期刊>Journal of bone and mineral metabolism >Changes in the expression of CD106, osteogenic genes, and transcription factors involved in the osteogenic differentiation of human bone marrow mesenchymal stem cells.
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Changes in the expression of CD106, osteogenic genes, and transcription factors involved in the osteogenic differentiation of human bone marrow mesenchymal stem cells.

机译:CD106,成骨基因和转录因子的表达变化涉及人骨髓间充质干细胞的成骨分化。

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Mesenchymal stem cells (MSCs) are well known to possess multipotential differentiation and are becoming a good tool for clinical research. However, specific markers for their purification and the mechanism of their osteogenic differentiation remain to be elucidated. In the present study, we compared the expression of CD106, and osteogenic differentiation-related proteins and genes in human bone marrow (BM)-derived MSCs, before and after differentiation by FACS, histochemical staining, immunohistochemical staining, RT-PCR, and real-time PCR. It was found that MSCs were positive for CD13, CD29, CD44, CD73, CD90, CD105, and CD166, but negative for CD14, CD31, CD34, CD62E, CD45, and GlyA. Notably, CD106 was detected before osteogenic induction, but its expression was downregulated 10 fold after 2 weeks of osteogenic differentiation as determined by flow cytometry. The results of RT-PCR and real-time PCR revealed that the expression of CD106 mRNA in MSCs significantly decreased by 7.1-, 4.2-, and 5.1-fold, respectively after osteogenic, chondrogenic, and adipogenic differentiation. In contrast, other MSC-positive markers described above did not change significantly even after differentiation. Compared to levels in control cells, after 2 weeks of osteogenic differentiation, mRNA levels of alkaline phosphatase, bone sialoprotein, osteocalcin, and transcript factors RUNX2 and Osterix showed more than 2-fold, 5-fold, 1.5-fold, 2-fold, and 5-fold increase, respectively. Thus, we speculate that CD106 might be a useful surface marker for BMMSCs. Moreover, alkaline phosphatase, type I collagen, osteonectin, osteopontin, and biglycin were expressed in the early stages of osteogenic differentiation before bone sialoprotein and osteocalcin. The present study should help to provide a novel marker for isolating purified MSCs and characterizing osteogenic differentiation.
机译:众所周知,间充质干细胞(MSC)具有多能分化能力,正在成为临床研究的良好工具。然而,其纯化的特定标记及其成骨分化的机制仍有待阐明。在本研究中,我们比较了通过FACS,组织化学染色,免疫组织化学染色,RT-PCR和真核细胞分化前后CD106以及成骨分化相关蛋白和基因在人骨髓(BM)MSC中的表达。实时PCR。发现MSC对CD13,CD29,CD44,CD73,CD90,CD105和CD166呈阳性,但对CD14,CD31,CD34,CD62E,CD45和GlyA呈阴性。值得注意的是,CD106在成骨诱导之前被检测到,但通过流式细胞术测定,其表达在成骨分化2周后被下调了10倍。 RT-PCR和实时PCR的结果表明,成骨,软骨形成和成脂分化后,MSCs中CD106 mRNA的表达分别显着降低了7.1、4.2和5.1倍。相反,即使分化后,上述其他MSC阳性标志物也没有明显变化。与对照组细胞相比,成骨分化2周后,碱性磷酸酶,骨唾液蛋白,骨钙素和转录因子RUNX2和Osterix的mRNA水平显示超过2倍,5倍,1.5倍,2倍,和5倍增加。因此,我们推测CD106可能是BMMSC有用的表面标记。此外,碱性磷酸酶,I型胶原蛋白,骨连接蛋白,骨桥蛋白和双糖蛋白在成骨细胞分化的早期就表达,然后是唾液酸蛋白和骨钙素。本研究应有助于提供一种新的标记物,用于分离纯化的MSC和表征成骨分化。

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