Optimization and Application of Liposome-Mediated Transfection of pEGFP-N1 in FRT Cells

Abstract

This study aimed to optimize liposome-mediatedtransfection conditions of pEGFP-N1 in FRT cells and to investigatewhether the optimized conditions were the optimal liposome-mediated transfectionconditions of the other vectors in FRT cells or not. The pEGFP-N1 were transfectedinto FRT cells, with the conditions of different cell confluence、ratio and quantity of vectors / liposome. At the sametime, pEGFP-N1-Aquaporin1、pEGFP-N1-Aquaporin3and pEGFP-N1-Aquaporin4 were transfected into FRT cells, respectively, withconditions of being same as pEGFP-N1. The inverted fluorescent microscopy was used to observe cytotoxicity and the expressionof EGFP in FRT cells. Transfection efficiency was measured by flow cytometry and cell viability was measured by trypanblue exclusion. The results showed that the expression of EGFP reachedthe highest at 36h after transfection. Flow cytometryand trypan blue exclusion tests showed when the cell confluence was 70%、the ratio and quantity of vectors / liposome was 1: 4(2.0ng: 8.0μL), pEGFP-N1 got higher transfection efficiency (46.97±0.32) % andcell viability (63.47±0.32) %. Under the same conditions, AQP1、AQP3 and AQP4 got the highest transfection efficiency and higher cell viability, too. The study would provide anexperimental evidence for efficient transfection of the other vectors in FRT cells.