Abstract
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/cm2, 1.0 W/cm2, 1.6 W/cm2, 2.2 W/cm2), 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/cm2, 2.2 W/cm2) 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/cm2 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.
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This work was supported by a grant from the National Natural Science Foundation of China (No. 30670548).
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Chen, Zy., Xie, Mx., Wang, Xf. et al. Different effects of therapeutic ultrasound parameters and culture conditions on gene transfection efficiency. Chin. J. Cancer Res. 20, 249–254 (2008). https://doi.org/10.1007/s11670-008-0249-z
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DOI: https://doi.org/10.1007/s11670-008-0249-z