Expression of OCT-4 and SOX-2 in Bone Marrow-Derived Human Mesenchymal Stem Cells during Osteogenic Differentiation.
Expression of OCT-4 and SOX-2 in Bone Marrow-Derived Human Mesenchymal Stem Cells during Osteogenic Differentiation.
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DOI：
10.3889/oamjms.2016.008
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发表时间：
2016-03-15
影响因子：
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通讯作者：
Marijanovic I
中科院分区：
文献类型：
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作者：
Matic I;Antunovic M;Brkic S;Josipovic P;Mihalic KC;Karlak I;Ivkovic A;Marijanovic I
测定多能性基因OCT-4和SOX-2在人间充质干细胞（hMSCs）成骨分化前后的表达水平。人骨髓间充质干细胞来源于骨髓并分化为成骨细胞。在细胞培养的第0天和第14天进行分析。由于骨标志物-碱性磷酸酶（AP）活性和AP和骨唾液蛋白（BSP）的信使RNA（mRNA）表达，评价了体外分化。OCT-4和SOX-2的表达通过实时qPCR在mRNA水平上进行评估，并通过免疫细胞化学在蛋白水平上进行评估。体外培养第14天显示AP活性增加，AP和BSP基因表达上调。第0天未分化hMSC中的OCT-4和SOX-2是可检测的，并且与作为阳性对照的肿瘤细胞系相比非常低。免疫细胞化学在分化前（第0天）和分化后（第14天）检测细胞核中的OCT-4。在相同的时间点，培养物对SOX-2蛋白呈阴性。从hMSC分离的多能性标志物OCT-4和SOX-2的信使RNA较少存在，而OCT-4蛋白在分化成骨细胞谱系之前和之后的细胞核中检测到。
Determine the levels of expression of pluripotency genes OCT-4 and SOX-2 before and after osteogenic differentiation of human mesenchymal stem cells (hMSCs). Human MSCs were derived from the bone marrow and differentiated into osteoblasts. The analyses were performed on days 0 and 14 of the cell culture. In vitro differentiation was evaluated due to bone markers – alkaline phosphatase (AP) activity and the messenger RNA (mRNA) expression of AP and bone sialoprotein (BSP). The OCT-4 and SOX-2 expression was evaluated at mRNA level by real-time qPCR and at protein level by immunocytochemistry. In vitro cultures on day 14 showed an increase in AP activity and upregulation of AP and BSP gene expression. OCT-4 and SOX-2 in undifferentiated hMSCs on day 0 is detectable and very low compared to tumor cell lines as a positive control. Immunocytochemistry detected OCT-4 in the cell nuclei prior (day 0) and post differentiation (day 14). On the same time points, cultures were negative for SOX-2 protein. Messenger RNA for pluripotency markers OCT-4 and SOX-2 isolated from hMSCs was less present, while OCT-4 protein was detected in cell nuclei prior and post differentiation into osteoblast lineage.