بهینه سازی و مقایسه سه حامل تجاری درترانسفکشن موثر به سلول‌های تک هسته ای خون محیطی (PBMCs)

Background & Aim: Peripheral Blood Mononuclear Cells (PBMCs) are the main components of the immune system, including lymphocytes and monocytes. These cells are being used in most immunological studies and therefore, isolation, culture and transfection of these cells are of great importance. The transfection of these cells is currently a key concern; this is because most common methods of transfection into the PBMCs are not well suited. The purpose of this study was to compare and optimize the effects of three commercial methods (PolyFect, Lipofectamine 2000 and Lipofectamine 3000) on PBMC transfection using PMAX-GFP vector. Methods: PBMC cells were isolated from the peripheral blood by Ficole.  Following the cell culture, the PMAX-GFP vector was transfected into the cells using the Polyfect, Lipopactamine 2000 and Lipopactamine 3000. After 24, 72 and 120 hours, the transfection rate for each method was evaluated by fluorescence microscope. Results: Based on the obtained results, the level of transfection for PBMCs by Lipopactamine 2000 and Lipopactamine 3000 at all times was about 2-3%. The amount of transfection of these cells by the Polyfect after 120 hours was about 70%. Conclusion: Generally, the efficiency of transfection of PBMCs s by Polyfect was probably the best in terms of increasing time, which introduces a new method for gene transfer into these cells. However, further studies are required to achieve the highest conversion efficiency of PBMC cells.