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Indirect co-culture with tenocytes promotes proliferation and mRNA expression of tendon/ligament related genes in rat bone marrow mesenchymal stem cells

机译:与肌腱细胞间接共培养可促进大鼠骨髓间充质干细胞的增殖和肌腱/韧带相关基因的mRNA表达

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Recent evidences have suggested that humoral factors released from the appropriate co-cultured cells influenced the expansion and differentiation of mesenchymal stem cells (MSCs). However, little is known about the proliferation and differentiation of MSCs subjected to co-culture condition with tenocytes. In this study, we aimed to establish a co-culture system of MSCs and tenocytes and investigate the proliferation and tendon/ligament related gene expression of MSCs. MTT assay was used to detect the expansion of MSCs. Semi-quantitative RT-PCR was performed to investigate the expression of proliferation associated c-fos gene and tendon/ligament related genes, including type I collagen (Col I), type III collagen (Col III), tenascin C and scleraxis. Significant increase in MSCs expansion was observed after 3 days of co-culture with tenocytes. The c-fos gene expression was found distinctly higher than for control group on day 4 and day 7 of co-culture. The mRNA expression of four tendon/ligament related genes was significantly up-regulated after 14 days of co-culture with tenocytes. Thus, our research indicates that indirect co-culture with tenocytes promotes the proliferation and mRNA expression of tendon/ligament related genes in MSCs, which suggests a directed differentiation of MSCs into tendon/ligament.
机译:最近的证据表明,从适当的共培养细胞释放的体液因子影响间充质干细胞(MSCs)的扩增和分化。然而,关于与肌腱细胞共培养的MSC的增殖和分化知之甚少。在这项研究中,我们旨在建立MSC和肌腱细胞的共培养系统,并研究MSC的增殖和与肌腱/韧带相关的基因表达。使用MTT测定法检测MSC的扩增。进行了半定量RT-PCR,以研究与增殖相关的c-fos基因和与肌腱/韧带相关的基因的表达,包括I型胶原(Col I),III型胶原(Col III),腱生蛋白C和硬化。与肌腱细胞共培养3天后,观察到MSCs的扩增显着增加。共培养第4天和第7天发现c-fos基因表达明显高于对照组。与肌腱细胞共培养14天后,四个与腱/韧带相关的基因的mRNA表达明显上调。因此,我们的研究表明,与肌腱细胞的间接共培养促进了MSC中肌腱/韧带相关基因的增殖和mRNA表达,这表明MSC定向分化为肌腱/韧带。

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