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Using cervical-carcinoma-derived cells as a model, the present study investigates the effects cell line and cell cycle phase have on sonoporation transfection efficiency under the same physical conditions. A plasmid expressing green fluorescent protein (GFP) was used to measure transfection efficiency. To evaluate the effect of cell type, CaSki, HeLa, and SiHa cells were sonoporated using an acoustic pressure of 1 MPa for 30 s with a duty cycle of 4.8% in the presence of the GFP plasmid. To study the effect of cell cycle phase, SiHa cells were synchronized at S-phase using a double thymidine block and sonoporated at different time points after the block. Contrast agent microbubbles were used at a 0.33% volume concentration. Results indicated that both cell line and cell cycle phase impact the transfection efficiency obtained with sonoporation.