Gonçalves Gabrielle V M, Silva Daniela N, Carvalho Rejane H, Souza Bruno S F, da Silva Kátia Nunes, Vasconcelos Juliana F, Paredes Bruno D, Nonaka Carolina K V, Ribeiro-Dos-Santos Ricardo, Soares Milena B P
Center for Biotechnology and Cell Therapy, Hospital São Rafael, Salvador, BA, 41253-190, Brazil.
Gonçalo Moniz Institute, Oswaldo Cruz Foundation (FIOCRUZ), Rua Waldemar Falcão, 121, Salvador, BA, 40296-710, Brazil.
Cytotechnology. 2018 Apr;70(2):577-591. doi: 10.1007/s10616-017-0131-2. Epub 2017 Sep 2.
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.
间充质干细胞（MSC）是细胞治疗和再生医学领域中很有前景的工具。除了其分化潜能外，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能会增强MSC的治疗效果。在此，我们生成并鉴定了过表达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生成了过表达hG-CSF或hIGF-1的MSC系。通过qRT-PCR对所产生克隆中hG-CSF或hIGF-1基因的表达进行定量，并通过ELISA在细胞上清液中检测蛋白质。这些细胞系表现出与对照MSC细胞系相似的细胞表面标志物以及向脂肪细胞、骨细胞和软骨细胞的分化潜能，表明即使经过基因修饰，它们的表型仍得以保留。IGF-1和G-CSF转基因细胞保持免疫抑制活性。最后，与对照细胞相比，我们通过qRT-PCR阵列对表达hIGF-1和hG-CSF的细胞系进行了比较基因表达分析。我们的结果表明，所产生的细胞系可能是细胞治疗的有用工具，适用于疾病模型中的测试。
