期刊
MOLECULAR MEDICINE REPORTS
卷 12, 期 3, 页码 4063-4070出版社
SPANDIDOS PUBL LTD
DOI: 10.3892/mmr.2015.4003
关键词
murine; mesenchymal stem cell; interleukin 1 receptor; antagonist; gene; lentivirus; transfection
资金
- Key Disciplines of Shanghai Municipal Education Commission [J50206]
- Shanghai Committee of Science and Technology, China [114119a2800]
- Chinese Academy of Sciences [XDA01030102]
- National Natural Science Foundation of China [81190133]
Genetically modified mesenchymal stein cells have been used in attempts to increase the expression of interleukin-1 receptor antagonist (IL-1Ra); however, the attempts thus far have been unsuccessful. The aim of the present study was to investigate whether the lentivirus transduced IL-1Ra gene was able to be stably expressed in murine bone marrow-derived mesenchymal stem cells (mBMSCs) in vitro. In the present study, third generation lentiviral (Lv) vectors transducing the IL-1Ra/green fluorescent protein (copGFP) gene were constructed and transfected into mBMSCs to establish the Lv-IL-1Ra.copGFP/mBMSCs, which were evaluated using fluorescence microscopy, flow cytometry, cell viability analysis using a cell counting kit-8 kit, Trypan blue staining and an MTT growth kinetics assay. The expression of IL-1Ra was analyzed using reverse transcription-quantitative polymerase chain reaction and western blotting. The results demonstrated that the Lv.IL-1Ra/copGFP vector was successfully constructed. The mBMSCs exhibited a short proliferation life, however they had good growth kinetics at an early stage and improved viability following efficient transduction of the IL-1Ra gene. IL-11Ra was overexpressed following transfection of fliBiNt.SCs. In conclusion, lentiviral vector transduced mBMSCs were able to efficiently express exogenous II-1Ra under certain conditions and had a marked capacity for proliferation.
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