目的观察慢病毒载体转染豚鼠巩膜成纤维细胞的有效性及对细胞活性的影响。方法转染组用慢病毒携带绿色荧光蛋白(lentiviral mediated green fluorescent protein,Lenti-GFP)转染豚鼠巩膜成纤维细胞,对照组加入空白培养液,转染组按最佳...目的观察慢病毒载体转染豚鼠巩膜成纤维细胞的有效性及对细胞活性的影响。方法转染组用慢病毒携带绿色荧光蛋白(lentiviral mediated green fluorescent protein,Lenti-GFP)转染豚鼠巩膜成纤维细胞,对照组加入空白培养液,转染组按最佳感染复数(multiplicity of infection,MOI)为50、100、200、400加入Lenti-GFP,转染2d、3d、4d、5d、6d和7d后,在倒置显微镜下观察绿色荧光蛋白表达情况,流式细胞仪检测转染效率,MTT法检测病毒对细胞增殖的影响。结果 Lenti-GFP转染细胞后48h可以看见绿色荧光;在同一MOI值下,随时间延长转染效率增高,第5天达到高峰;在MOI为400,转染5d时转染效率达82.86%;转染5d行MTT检测显示:MOI为50～400时,同一转染时间各组IOD值与对照组比较,差异均无统计学意义(均为P>0.05)。结论慢病毒载体可以在体外稳定而有效地转染豚鼠巩膜成纤维细胞,且不影响细胞活性,是巩膜成纤维细胞理想的基因转染载体。展开更多
In this paper, we report a novel method to synthesize polyethylenimine (PEI) nanogels in the range of 80— 200nm with narrow size distribution by photo-chemistry at room temperature in aqueous solution. The nanogels’...In this paper, we report a novel method to synthesize polyethylenimine (PEI) nanogels in the range of 80— 200nm with narrow size distribution by photo-chemistry at room temperature in aqueous solution. The nanogels’ size, size distributions and zeta potential were determined by photo correlation spectroscopy (PCS). Spherical morphology of the nanogels was characterized by scanning electron microscopy (SEM) and confirmed by atomic force microscopy (AFM). The nanogels are of high stability, low toxicity and low immunogenicity, as having been confirmed in in vivo tests with mice as animal model, and in vitro tests with human lung and liver cancer cells as well. Efforts are being made for further studies on synthesis and applications of the nanogels.展开更多
文摘目的观察慢病毒载体转染豚鼠巩膜成纤维细胞的有效性及对细胞活性的影响。方法转染组用慢病毒携带绿色荧光蛋白(lentiviral mediated green fluorescent protein,Lenti-GFP)转染豚鼠巩膜成纤维细胞,对照组加入空白培养液,转染组按最佳感染复数(multiplicity of infection,MOI)为50、100、200、400加入Lenti-GFP,转染2d、3d、4d、5d、6d和7d后,在倒置显微镜下观察绿色荧光蛋白表达情况,流式细胞仪检测转染效率,MTT法检测病毒对细胞增殖的影响。结果 Lenti-GFP转染细胞后48h可以看见绿色荧光;在同一MOI值下,随时间延长转染效率增高,第5天达到高峰;在MOI为400,转染5d时转染效率达82.86%;转染5d行MTT检测显示:MOI为50～400时,同一转染时间各组IOD值与对照组比较,差异均无统计学意义(均为P>0.05)。结论慢病毒载体可以在体外稳定而有效地转染豚鼠巩膜成纤维细胞,且不影响细胞活性,是巩膜成纤维细胞理想的基因转染载体。
文摘In this paper, we report a novel method to synthesize polyethylenimine (PEI) nanogels in the range of 80— 200nm with narrow size distribution by photo-chemistry at room temperature in aqueous solution. The nanogels’ size, size distributions and zeta potential were determined by photo correlation spectroscopy (PCS). Spherical morphology of the nanogels was characterized by scanning electron microscopy (SEM) and confirmed by atomic force microscopy (AFM). The nanogels are of high stability, low toxicity and low immunogenicity, as having been confirmed in in vivo tests with mice as animal model, and in vitro tests with human lung and liver cancer cells as well. Efforts are being made for further studies on synthesis and applications of the nanogels.
