Department of Emergency Orthopedics, Affiliated Hospital of Guizhou Medical University, China.
J Cell Biochem. 2018 Nov;119(10):8389-8397. doi: 10.1002/jcb.27034. Epub 2018 Jul 3.
To investigate the effect of lentiviral transfection containing bFGF gene on the biological characteristics of bone marrow stromal cells (BMSCs). BMSCs were obtained by density gradient centrifugation and adherence screening. The bFGF gene was transfected into BMSCs by lentiviral vector and divided into bFGF transfection group, empty virus group, and untransfected group according to the transfection conditions. After transfection, the morphology, expression of bFGF mRNA and protein, cell proliferation, cell cycle, and alkaline phosphatase (ALP) activity were observed in three groups of cells. High density BMSCs were successfully obtained by density gradient centrifugation and adherence screening. After transfection of BMSCs with bFGF gene, the cell morphology did not change significantly, while the expression of bFGF mRNA and protein were significantly increased, the cell proliferation curve shifted upward, the proportion of proliferating cells increased, and the activity of ALP was significantly enhanced. Compared with empty virus group and untransfected group, there was significant difference (P < 0.05). The rabbit bFGF gene was successfully introduced into the BMSCs cultured in vitro by lentiviral vector, and the target gene was stably expressed. And the expression of bFGF can promote the proliferation and osteogenic differentiation of BMSCs.
探讨携带碱性成纤维细胞生长因子(bFGF)基因的慢病毒转染对骨髓基质细胞(BMSCs)生物学特性的影响。
采用密度梯度离心和贴壁筛选法分离培养 BMSCs,通过慢病毒载体将 bFGF 基因转染至 BMSCs,根据转染条件将其分为 bFGF 转染组、空病毒组和未转染组。观察三组细胞的形态、bFGFmRNA 及蛋白表达、细胞增殖、细胞周期及碱性磷酸酶(ALP)活性。
成功通过密度梯度离心和贴壁筛选获得高浓度 BMSCs。bFGF 基因转染 BMSCs 后,细胞形态无明显变化,bFGFmRNA 及蛋白表达明显增加,细胞增殖曲线向上偏移,增殖细胞比例增加,ALP 活性明显增强,与空病毒组和未转染组比较,差异均有统计学意义(P<0.05)。
成功通过慢病毒载体将兔 bFGF 基因导入体外培养的 BMSCs 中,并稳定表达目的基因,且 bFGF 的表达可促进 BMSCs 的增殖和向成骨细胞分化。