Eslami Asma, Dehbashi Moein, Ashja-Arvan Mehnoosh, Salehi Hossein, Azimzadeh Maryam, Ganjalikhani-Hakemi Mazdak
Department of Immunology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran.
Division of Genetics, Department of Cell and Molecular Biology, Faculty of Biological Sciences and Technologies, University of Isfahan, 81746-73441, Isfahan, Iran.
Cytotechnology. 2020 Oct;72(5):773-784. doi: 10.1007/s10616-020-00421-8. Epub 2020 Sep 15.
Adipose-derived stem cells (ADSCs) are a type of mesenchymal stem cells with the therapeutic effects that make them one of the best sources for cell therapy. In this study, we aimed to assess the ability of human ADSCs for constant expression of IL-11 and IL-13, simultaneously. In this study, the characterized hADSCs were transduced with a lentiviral vector (PCDH-513B) containing IL-11 and IL-13 genes, and the ability of long-term expression of the transgenes was evaluated by ELISA technique on days 15, 45 and 75 after transduction. Our results indicated a high rate of transduction (more than 90%) in the isolated hADSCs. Our data showed the highest rate of expression on days 75 after transduction which was 242.67 pg/ml for IL-11 and 303.6 pg/ml for IL-13 compared with 35.2 pg/ml and 35.6 pg/ml in untreated cells, respectively (p = 0.001). Besides, MTT assay showed transduction of hADSCs with lentiviral viruses containing IL-11 and IL-13 had no adverse effect on hADSCs proliferation (p-value = 0.89). Finally, we successfully constructed a hADSC population stably overexpressing IL-11 as the neurotrophic cytokine and IL-13 as the anti-inflammatory cytokine and this transduced cells can be used for further studies in EAE mice model.
脂肪来源干细胞（ADSCs）是一种间充质干细胞，具有治疗作用，使其成为细胞治疗的最佳来源之一。在本研究中，我们旨在评估人ADSCs同时持续表达IL-11和IL-13的能力。在本研究中，用含有IL-11和IL-13基因的慢病毒载体（PCDH-513B）转导已鉴定的hADSCs，并在转导后第15、45和75天通过ELISA技术评估转基因的长期表达能力。我们的结果表明，分离的hADSCs转导率很高（超过90%）。我们的数据显示，转导后第75天表达率最高，IL-11为242.67 pg/ml，IL-13为303.6 pg/ml，而未处理细胞中分别为35.2 pg/ml和35.6 pg/ml（p = 0.001）。此外，MTT试验表明，用含有IL-11和IL-13的慢病毒转导hADSCs对hADSCs增殖没有不良影响（p值 = 0.89）。最后，我们成功构建了一个稳定过表达作为神经营养细胞因子的IL-11和作为抗炎细胞因子的IL-13的hADSC群体，这种转导细胞可用于在实验性自身免疫性脑脊髓炎（EAE）小鼠模型中进行进一步研究。
