Interrelation between HeLa-S3 cell transfection and hemolysis in red blood cell suspension using pulsed ultrasound of various duty cycles |
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Authors: | Y Liu H Uno H Takatsuki M Hirano A Sakanishi |
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Institution: | (1) Department of Biological and Chemical Engineering, Faculty of Engineering, Gunma University, Kiryu, Gunma 376-8515, Japan |
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Abstract: | We have studied the in vitro transfection of a plasmid DNA with the lacZ gene to HeLa-S3 cells and hemolysis in a red blood cell (RBC) suspension under pulsed ultrasound with duty cycles of 10, 20 and 30% using a digital sonifier at a frequency of 20 kHz and an intensity of 6.2 W/cm2 on the surface of a horn tip. Cultured HeLa-S3 cells in suspension were exposed to pulsed ultrasound for an apparent exposure time t from 0 to 60 s. HeLa-S3 viability decreased as a single exponential function of the total exposure time t= t with a common time constant =3.8 s for three duty cycles. Transfection was evaluated by counting the number of -galactosidase( -Gal)-positive cells relative to the total number of cells. Pulsed ultrasound provided an enhanced transfer of the -Gal plasmid to HeLa-S3 cells, 3.4-fold as compared with that in the case of the control. The optimal transfection efficiencies were 0.75, 0.80 and 0.74% near t= with =10, 20 and 30%, respectively. The number ratio of -Gal-positive cells to the surviving cells after exposure increased with t according to a modified logistic equation. The degree of hemolysis also increased exponentially with t at a time constant ![tau](/content/fnh5gvcebmvnpuup/xxlarge964.gif) = 0/ for the RBC suspension in physiological saline at a hematocrit concentration of 0.5% with 0=0.9 s. Thus the total exposure time for the optimal transfection efficiency was , that is, nearly four times of 0. Hemolysis in the RBC suspension may be a useful model for determining optimal transfection by pulsed ultrasound of various duty cycles. |
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Keywords: | Gene transfer HeLa-S3 cells Logistic equation Pulsed ultrasound Red blood cells |
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