水压转染法在小鼠不同肝叶的表达效率

The Different Efficiency of Hydrodynamics-based Transfection in Mouse Liver Lobes

为了解水压转染法(hydrodynamics-based transfection,HDT)在小鼠肝脏不同肝叶的转染效率,将容量为小鼠体重的10%,绿色荧光蛋白基因质粒pEGFP-C1含量为35μg/只的生理盐水溶液以0.4 mL/s的速度从小鼠尾静脉注射,于注射后不同时间取小鼠各肝叶制备冰冻切片,在荧光显微镜下观察,计数各肝叶的绿色荧光蛋白表达情况。结果显示注射后12 h,绿色荧光蛋白阳性细胞比例最高,从24 h起表达量逐渐减少,至48 h时各肝叶均基本难以检测出绿色荧光蛋白阳性细胞。在12 h观察各肝叶的转染效率如下:右叶、蒂状叶的绿色荧光蛋白阳性细胞约为22%,左叶、中叶、尾状叶约为15%取材部位不同会造成数据分析的极显著差异。

To understanding the efficiency of hydrodynamics-based transfection method in mouse different liver lobes, we inject capacity of mouse weight 10%, content 35 ug plasmid （pEGFP-C1, a kind of green fluorescent plasmid） each, and inject them with 0.5 mL/s, then collect mice＇ liver lobes at different time points after injection to make frozen section, finally observe and quantify the GFP expression with fluorescence microscope and count the expression of each lobe of liver. When one-off injection, the optimal conditions is that, at 12 h after injection, GFP- positive cells take on the highest proportion of above 20%; GFP begin to gradually reduce since 24 h, until 48 h almost no GFP-positive cells are seen in any liver lobe; briefly, under the above optimal condition, GFP expression in each liver lobe is described as follows： about 22% for right and pedicle lobe, about 15% for left, middle and caudate lobes, different part of materials can get very significant difference in transfection efficiency.