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Effects of human epidermal growth factor gene-transfected mesenchymal stem cells on fibroblast migration and proliferation

机译:人表皮生长因子基因转染的间充质干细胞对成纤维细胞迁移和增殖的影响

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Objectives: We were interested in determining whether epidermal growth factor gene-transfected mesenchymal stem cells (EGF-MSC) would accelerate fibroblast migration and proliferation. Materials and methods: Fibroblasts were cultured in serum-free conditioned media from EGF-MSC; RT-PCR was performed to detect expression of EGF gene in EGF-MSCs. EGF protein levels in cell culture supernatants from EGF-MSC were assayed by ELISA and proliferation of EGF-MSC-treated fibroblasts was performed using MTT assay. Effects of EGF-MSC on fibroblast migration were evaluated using scratch wound and transmigration assays. Cell adhesion molecules, cell dynamics molecules and phospho-(Ser) kinase substrate expressions of EGF-MSC-treated fibroblasts were evaluated by western blotting. Results: EGF gene expression increased in EGF-MSCs and viability of EGF-MSC-treated fibroblasts was elevated. EGF-MSC-treated fibroblasts showed increased migration compared to controls. Expressions of cell adhesion molecules (β-catenin, N-cadherin), cell dynamics molecules (cofilin, ezrin) and phospho-(Ser) kinase substrates (phospho-MAPK/CDK substrate, phospho-Arg-(Ser)-X-Tyr/Phe-X-pSer motif) increased in EGF-MSC-treated fibroblasts. These results imply that EGF-MSCs contributed to enhancing the wound healing process by increased cell adhesion, dynamic effects, fibroblast migration, and proliferation. Conclusions: This study indicates that EGF-MSCs had a positive influence on fibroblast migration and proliferation and EGF-MSC may provide a useful strategy for wound healing.
机译:目的:我们感兴趣的是确定表皮生长因子基因转染的间充质干细胞(EGF-MSC)是否会加速成纤维细胞的迁移和增殖。材料与方法:成纤维细胞在EGF-MSC的无血清条件培养基中培养。进行RT-PCR以检测EGF-MSC中EGF基因的表达。通过ELISA测定来自EGF-MSC的细胞培养上清液中的EGF蛋白水平,并使用MTT测定法进行经EGF-MSC处理的成纤维细胞的增殖。 EGF-MSC对成纤维细胞迁移的影响使用刮擦伤口和迁移测定法进行评估。通过蛋白质印迹法评估了EGF-MSC处理的成纤维细胞的细胞粘附分子,细胞动力学分子和磷酸-(Ser)激酶底物表达。结果:EGF-MSCs中EGF基因表达增加,EGF-MSC处理的成纤维细胞活力提高。与对照相比,EGF-MSC处理的成纤维细胞显示出增加的迁移。细胞粘附分子(β-catenin,N-钙黏着蛋白),细胞动力学分子(cofilin,ezrin)和磷酸-(Ser)激酶底物(磷酸-MAPK / CDK底物,磷酸-Arg-(Ser)-X-Tyr)的表达/ Phe-X-pSer基序)在EGF-MSC处理的成纤维细胞中增加。这些结果暗示,EGF-MSC通过增加细胞粘附,动力作用,成纤维细胞迁移和增殖来促进伤口愈合过程。结论:该研究表明EGF-MSCs对成纤维细胞的迁移和增殖具有积极的影响,EGF-MSCs可能为伤口愈合提供有用的策略。

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