家畜生态学报2024，Vol.45Issue(2)：13-18,6.DOI:10.3969/j.issn.1673-1182.2024.02.003
不同转染方法应用于体细胞转基因效率的研究
Comparative Study of the Efficiency of Different Methods Transfected Somatic Cells
摘要
Abstract
The objective of this paper is to compare the effect of different transfection methods media-ting the transfection of enhanced green fluorescent protein(EGFP)gene into different porcine somatic cells.Porcine adipose and bone marrow-derived mesenchymal stem cells(MSCs)and porcine fetal fibro-blasts were isolated and cultured.The plasmid containing EGFP was transfected into each of the three cell types using electroporation,liposome and Roche transfection reagent,respectively.Fluorescence expres-sion was observed by inverted fluorescence microscopy,and relative cell viability and transfection efficiency were analyzed by blood count and flow cytometry.The results showed that EGFP expression was observed in all three methods of transfection of different somatic cells for 48 hours.In the comparison of different methods of transfecting the same somatic cells,the cell survival rate was in the order of Roche transfection reagent＞liposome＞electroporation method,the survival rate of cells transfected with Roche transfec-tion reagent and liposome was significantly higher than that of electroporation(P＜0.05).The transfection efficiency in descending order was electroporation＞Roche transfection reagent＞liposome,with electro-poration significantly higher than Roche transfection reagent and liposome,the transfection efficiency of Roche transfection reagent was also significantly higher than that of liposome(P＜0.05).In the compari-son of transfection of different somatic cells using the same method,there was no significant difference in the survival rate of the three somatic cells after transfection by the same method(P＞0.05),and the transfection efficiency of MSCs by electroporation were significantly higher than the porcine fetal fibro-blasts(P＜0.05).There was no significant difference in transfection efficiency between the three types of somatic cells transfected with Roche transfection reagent and liposomes respectively(P＞0.05).All three methods can be used to transfect porcine somatic cells.Considering transfection efficiency and cell survival,the electroporation method is the most effective and can be used to transfect cells in good condition,fol-lowed by Roche reagent transfection.关键词
电穿孔法/脂质体/罗氏转染试剂/体细胞/转染效率/细胞存活率Key words
electroporation/liposome/Roche transfection reagent/somatic cells/transfection efficien-cy/cell viability分类
农业科技引用本文复制引用
赵广伟,杨小玲,张孟奇,苏勤智,陈凤..不同转染方法应用于体细胞转基因效率的研究[J].家畜生态学报,2024,45(2):13-18,6.基金项目
河南牧业经济学院博士科研启动基金项目(24030028) （24030028）
