Ultrasound Transfection A Gene Transfer Assay for Increased Cell Permeability without Impacting Cell Viability
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Authors
Anderson, Brianne
Date of Issue
2000-04-01
Type
thesis
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Abstract
In this study, NIH/3T3 cells were permeated with both chemical and physical methods for transient gene transfection. Cationic lipid transfection was used as the mode of chemical transfection with Transfectam used as the transfection reagent and pSV-P- galactosidase as the control vector. After addition of transfection agents, cells were exposed to ultrasound with either of two ultrasound probe-type sonicators: either a 20 kHz, 130 W Vibra-Cell Ultrasonic Processor or a 20 kHz, 400 W Vibra-Cell Ultrasonic Processor. With the 130 W Vibra-Cell Ultrasonic Processor, no correlation could be made between transfection rates and ultrasound exposure (duration or power). Using the 400 W Vibra-Cell Ultrasonic Processor, it was determined that a 1% amplitude for 30 seconds may represent the optimal set of conditions to enhance cell permeability and that the viability of the cells decreases at higher levels of amplitude.