{"title":"Comparing chemical transfection, electroporation, and lentiviral vector transduction to achieve optimal transfection conditions in the Vero cell line.","authors":"Parisa Jamour, Abbas Jamali, Arash Ghalyanchi Langeroudi, Behrouz Ebadi Sharafabad, Asghar Abdoli","doi":"10.1186/s12860-024-00511-x","DOIUrl":null,"url":null,"abstract":"<p><strong>Background: </strong>Transfection is an important analytical method for studying gene expression in the cellular environment. There are some barriers to efficient DNA transfection in host cells, including circumventing the plasma membrane, escaping endosomal compartmentalization, autophagy, immune sensing pathways, and translocating the nuclear envelope. Therefore, it would be very useful to introduce an optimum transfection approach to achieve a high transfection efficiency in the Vero cell line. The aim of this study was to compare various transfection techniques and introduce a highly efficient method for gene delivery in Vero cells.</p><p><strong>Methods: </strong>In the current study, three transfection methods were used, including chemical transfection, electroporation, and lentiviral vector transduction, to obtain the optimum transfection conditions in the Vero cell line. Vero cells were cultured and transfected with chemical transfection reagents, electroporation, or HIV-1-based lentivectors under different experimental conditions. Transfection efficiency was assessed using flow cytometry and fluorescence microscopy to detect GFP-positive cells.</p><p><strong>Results: </strong>Among the tested methods, TurboFect™ chemical transfection exhibited the highest efficiency. Optimal transfection conditions were achieved using 1 µg DNA and 4 µL TurboFect™ in 6 × 10<sup>4</sup> Vero cells.</p><p><strong>Conclusion: </strong>TurboFect™, a cationic polymer transfection reagent, demonstrated superior transfection efficiency in Vero cells compared with electroporation and lentivirus particles, and is the optimal choice for chemical transfection in the Vero cell line.</p>","PeriodicalId":2,"journal":{"name":"ACS Applied Bio Materials","volume":null,"pages":null},"PeriodicalIF":4.6000,"publicationDate":"2024-05-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11089686/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"ACS Applied Bio Materials","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1186/s12860-024-00511-x","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"MATERIALS SCIENCE, BIOMATERIALS","Score":null,"Total":0}
引用次数: 0
Abstract
Background: Transfection is an important analytical method for studying gene expression in the cellular environment. There are some barriers to efficient DNA transfection in host cells, including circumventing the plasma membrane, escaping endosomal compartmentalization, autophagy, immune sensing pathways, and translocating the nuclear envelope. Therefore, it would be very useful to introduce an optimum transfection approach to achieve a high transfection efficiency in the Vero cell line. The aim of this study was to compare various transfection techniques and introduce a highly efficient method for gene delivery in Vero cells.
Methods: In the current study, three transfection methods were used, including chemical transfection, electroporation, and lentiviral vector transduction, to obtain the optimum transfection conditions in the Vero cell line. Vero cells were cultured and transfected with chemical transfection reagents, electroporation, or HIV-1-based lentivectors under different experimental conditions. Transfection efficiency was assessed using flow cytometry and fluorescence microscopy to detect GFP-positive cells.
Results: Among the tested methods, TurboFect™ chemical transfection exhibited the highest efficiency. Optimal transfection conditions were achieved using 1 µg DNA and 4 µL TurboFect™ in 6 × 104 Vero cells.
Conclusion: TurboFect™, a cationic polymer transfection reagent, demonstrated superior transfection efficiency in Vero cells compared with electroporation and lentivirus particles, and is the optimal choice for chemical transfection in the Vero cell line.
背景:转染是研究细胞环境中基因表达的重要分析方法:转染是研究细胞环境中基因表达的一种重要分析方法。在宿主细胞中高效转染 DNA 存在一些障碍,包括绕过质膜、逃脱内体分隔、自噬、免疫传感途径和转运核膜。因此,引入一种最佳转染方法来实现 Vero 细胞系的高转染效率是非常有用的。本研究旨在比较各种转染技术,并引入一种在 Vero 细胞中高效传递基因的方法:本研究采用了三种转染方法,包括化学转染、电穿孔和慢病毒载体转导,以获得 Vero 细胞系的最佳转染条件。在不同的实验条件下,培养 Vero 细胞并用化学转染试剂、电穿孔或基于 HIV-1 的慢病毒载体进行转染。使用流式细胞术和荧光显微镜检测 GFP 阳性细胞,评估转染效率:结果:在所有测试方法中,TurboFect™ 化学转染的效率最高。在 6 × 104 个 Vero 细胞中使用 1 µg DNA 和 4 µL TurboFect™ 可达到最佳转染条件:结论:与电穿孔和慢病毒颗粒相比,阳离子聚合物转染试剂 TurboFect™ 在 Vero 细胞中的转染效率更高,是 Vero 细胞系化学转染的最佳选择。