Generation and characterization of transgenic mouse mesenchymal stem cell lines expressing hIGF-1 or hG-CSF
Generation and characterization of transgenic mouse mesenchymal stem cell lines expressing hIGF-1 or hG-CSF
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DOI：
10.1007/s10616-017-0131-2
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发表时间：
2018-04-01
期刊：
影响因子：
2.2
通讯作者：
Soares, Milena B. P.
中科院分区：
文献类型：
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作者：
Goncalves, Gabrielle V. M.;Silva, Daniela N.;Soares, Milena B. P.
间充质干细胞（MSC）是细胞治疗和再生医学领域的有前途的工具。除了它们的分化潜力，MSC还具有分泌刺激组织再生的生物活性分子的能力。因此，细胞因子和生长因子的过表达可增强MSC的治疗效果。在这里，我们产生并表征了过表达hG-CSF或hIGF-1的小鼠骨髓MSC系。通过慢病毒载体介导的基因转移产生过表达hG-CSF或hIGF-1的MSC系。通过qRT-PCR定量产生的克隆中hG-CSF或hIGF-1基因的表达，并通过ELISA检测细胞上清液中的蛋白质。细胞系显示细胞表面标志物和分化成脂肪细胞、骨细胞和软骨细胞的潜力，与对照MSC细胞系相似，表明即使在遗传修饰后其表型也保持不变。IGF-1和G-CSF转基因细胞保持免疫抑制活性。最后，当与对照细胞相比时，我们通过qRT-PCR阵列在表达hIGF-1和hG-CSF的细胞系中进行比较基因表达分析。我们的研究结果表明，所产生的细胞系可能是细胞治疗的有用工具，适合在疾病模型中进行测试。
Mesenchymal stem cells (MSC) are promising tools in the fields of cell therapy and regenerative medicine. In addition to their differentiation potential, MSC have the ability to secrete bioactive molecules that stimulate tissue regeneration. Thus, the overexpression of cytokines and growth factors may enhance the therapeutic effects of MSC. Here we generated and characterized mouse bone marrow MSC lines overexpressing hG-CSF or hIGF-1. MSC lines overexpressing hG-CSF or hIGF-1 were generated through lentiviral vector mediated gene transfer. The expression of hG-CSF or hIGF-1 genes in the clones produced was quantified by qRT-PCR, and the proteins were detected in the cell supernatants by ELISA. The cell lines displayed cell surface markers and differentiation potential into adipocytes, osteocytes and chondrocytes similar to the control MSC cell lines, indicating the conservation of their phenotype even after genetic modification. IGF-1 and G-CSF transgenic cells maintained immunosuppressive activity. Finally, we performed a comparative gene expression analysis by qRT-PCR array in the cell lines expressing hIGF-1 and hG-CSF when compared to the control cells. Our results demonstrate that the cell lines generated may be useful tools for cell therapy and are suitable for testing in disease models.