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摘要
Objective: To investigate the effect of different therapeutic ultrasound (TUS) parameters and culture conditions on the cell viability and transfection efficiency of human cervical cancer cells ( HeLa). Methods: HeLa cells were cultured using two different protocols ( in suspension or in monolayer). Subsequently, cells were exposed to different TUS intensity (0.4 W/cm(2), 1.0 W/cm(2), 1.6 W/cm(2), 2.2 W/cm(2)), duty cycle (DC)(10%, 20%, 50%), exposure time ( 1 min or 3 min). Cell viability was analyzed by flow cytometry. Gene transfection of red fluorescent protein (DsRED) was detected. Results: TUS intensity and duty cycle had a great impact on the overall results (P < 0.01). Cell injury were found to increase progressively with intensity (1.6 W/ cm(2), 2.2 W/cm(2)) and duty cycle (50%) and cell detachment was accompanied by ultrasound exposure in adherent cells. Results of factorial design showed that the fashion of cell culture and the TUS parameters had interaction (P < 0.01). The ideal conditions that cell viability above 80% producing maximum efficiency were noted to be at 1.0 W/cm(2) irradiated 3 min with a duty cycle of 20% in cell suspension. Conclusion: TUS parameters and transfection conditions have a great impact on the gene transfection and cell viability. Optimal parameters could enhance cell membrane permeability, which facilitate to delivering the macromolecules into cells.
- 出版日期2008-12
- 单位华中科技大学
