Optimization of Electrotransfection Conditions of BGC823 Cells

Abstract

Objectives: The GFP plasmid was transfected into BGC823 cells by electroporation, and the transfection conditions were optimized to obtain a high transfection rate. Methods: In the 400 μL electrotransfection system, BGC823 cells were transfected with different electric field intensity (150 V, 175 V, 200 V and 225 V), pulse time (10 ms, 30 ms, 50 ms and 70 ms), electric field intensity and pulse time (150 V, 70 ms; 175 V, 50 ms; 200 V, 30 ms; 225 V, 10 ms), and electrotransfection buffer solution (serum-free 1640, complete medium, Hepes buffer solution, PBS buffer solution and D-Hanks solution), and the survival rate and transfection rate of cells under different conditions were detected. Results: When the electric field intensity was 175 V, the pulse time was 30 ms, and the electrotransfection buffer solution was serum-free 1640, the better electrotransfection efficiency was achieved, with a survival rate of (37.35 ± 1.85)% and a transfection rate of (74.79 ± 1.89)%. Conclusions: The electrotransfection conditions of human gastric cancer BGC823 cells are optimized and the efficiency of electrotransfection is improved on the premise of guaranteeing the relatively high survival rate of cells, which lays the foundation for subsequent studies on the expression of recombinant proteins and antibodies, as well as gene therapy for gastric cancer cells.