背景与目的:选择合适的载体是基因治疗成功的关键之一,近年来一种新型多聚阳离子化合物——聚乙烯亚胺(polyethylenimine,PEI)作为高效低毒的基因转导载体得到广泛重视。本研究应用光化学法制备系列不同粒径的PEI纳米凝胶,初步探讨其转...背景与目的:选择合适的载体是基因治疗成功的关键之一,近年来一种新型多聚阳离子化合物——聚乙烯亚胺(polyethylenimine,PEI)作为高效低毒的基因转导载体得到广泛重视。本研究应用光化学法制备系列不同粒径的PEI纳米凝胶,初步探讨其转导效率与粒径之间的关系,筛选理想的肿瘤基因转导载体。方法:光化学法制备PEI,光相干光谱仪测定粒径,并用扫描电子显微镜和原子力显微镜表征;以PEI为载体将增强型绿色荧光蛋白(enhanced green fluorescence protein,EGFP)报告基因分别转入Bel7402细胞和A549细胞,荧光显微镜下计数和流式细胞仪检测转染率。结果:光相干光谱仪检测PEI粒径38～200nm,扫描电子显微镜和原子力显微镜检测其形貌多为球形。荧光显微镜下计数和流式细胞仪检测86.9nmPEI(4μg)介导EGFP基因(2μg)时转染率最高,Bel7402细胞分别为(32.75±1.01)%、(32.40±1.41)%,A549细胞分别为(29.81±1.84)%、(30.00±1.86)%;与脂质体相比差异无统计学意义(P>0.05)。结论:光化学法合成的PEI是有效的基因转导载体,PEI粒径为86.9nm转染最为有效。展开更多
Phospholipids are believed to be important biomaterials. However, limited information is available on their cytocompatibilities. The objective of this study is to evaluate the effects of different phospholipids on the...Phospholipids are believed to be important biomaterials. However, limited information is available on their cytocompatibilities. The objective of this study is to evaluate the effects of different phospholipids on the proliferation of hepatic Bel\|7402 cells by comparing the adhesion, viability and proliferation of Bel\|7402 cells cultured on different phospholipid surfaces. The cell adhesion, determined by counting the number of adhered cells to the surface, indicated that the cell adhesion was enhanced on charged phospolipid membranes. The cell viability evaluated by MTT\[3\|(4,5\|dimethylthiazole\|2\|yl)\|2,5\|diphenyl tetrazolium\|bromide\] showed that cells cultured on charged phospholipids have greater viability than those cultured on the control, while cells cultured on neutral phospholipids showed lower viability. The cell cycle analysis using flow cytometry demonstrated that S phase entry increased on charged phospholipids, while S phase entry decreased on neutral phospholipids. The results suggested that charged phospholipids, especially positively charged phospholipids, show better cytocompatibilities than neutral phospholipids to hepatic Bel\|7402 cell.展开更多
文摘背景与目的:选择合适的载体是基因治疗成功的关键之一,近年来一种新型多聚阳离子化合物——聚乙烯亚胺(polyethylenimine,PEI)作为高效低毒的基因转导载体得到广泛重视。本研究应用光化学法制备系列不同粒径的PEI纳米凝胶,初步探讨其转导效率与粒径之间的关系,筛选理想的肿瘤基因转导载体。方法:光化学法制备PEI,光相干光谱仪测定粒径,并用扫描电子显微镜和原子力显微镜表征;以PEI为载体将增强型绿色荧光蛋白(enhanced green fluorescence protein,EGFP)报告基因分别转入Bel7402细胞和A549细胞,荧光显微镜下计数和流式细胞仪检测转染率。结果:光相干光谱仪检测PEI粒径38～200nm,扫描电子显微镜和原子力显微镜检测其形貌多为球形。荧光显微镜下计数和流式细胞仪检测86.9nmPEI(4μg)介导EGFP基因(2μg)时转染率最高,Bel7402细胞分别为(32.75±1.01)%、(32.40±1.41)%,A549细胞分别为(29.81±1.84)%、(30.00±1.86)%;与脂质体相比差异无统计学意义(P>0.05)。结论:光化学法合成的PEI是有效的基因转导载体,PEI粒径为86.9nm转染最为有效。
基金theNationalNaturalScienceFoundationof China! (No .5 94932 0 8)
文摘Phospholipids are believed to be important biomaterials. However, limited information is available on their cytocompatibilities. The objective of this study is to evaluate the effects of different phospholipids on the proliferation of hepatic Bel\|7402 cells by comparing the adhesion, viability and proliferation of Bel\|7402 cells cultured on different phospholipid surfaces. The cell adhesion, determined by counting the number of adhered cells to the surface, indicated that the cell adhesion was enhanced on charged phospolipid membranes. The cell viability evaluated by MTT\[3\|(4,5\|dimethylthiazole\|2\|yl)\|2,5\|diphenyl tetrazolium\|bromide\] showed that cells cultured on charged phospholipids have greater viability than those cultured on the control, while cells cultured on neutral phospholipids showed lower viability. The cell cycle analysis using flow cytometry demonstrated that S phase entry increased on charged phospholipids, while S phase entry decreased on neutral phospholipids. The results suggested that charged phospholipids, especially positively charged phospholipids, show better cytocompatibilities than neutral phospholipids to hepatic Bel\|7402 cell.
