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A Novel Approach for Introducing Bio-Materials Into Cells
Authors:Chen  Da-Ren  Wendt  Christine H  Pui  David YH
Institution:(1) Particle Technology Laboratory, Department of Mechanical Engineering, University of Minnesota, 111 Church Street S.E., Minneapolis, MN 55455, USA;(2) Pulmonary and Critical Care, Department of Medicine, Medical School, University of Minnesota, 420 Delaware Street S.E., Minneapolis, MN 55455, USA;(3) Particle Technology Laboratory, Department of Mechanical Engineering, University of Minnesota, 111 Church Street S.E., Minneapolis, MN 55455, USA
Abstract:A novel approach was developed to introduce biological materials into cells for gene transfection and gene therapy applications. The method is based on the technique of electrospraying bio-materials into cells. A prototype apparatus was constructed for a feasibility study. The features of the gene transfector include: (1) A dual-capillary assembly to spray suspensions of biological materials. The outer capillary provided sheathing liquid that controlled the charge level on individual particles without altering the properties of suspensions. (2) An air–CO2 gas mixture was used for suppressing possible corona discharge and kept the same gas composition as those in incubators. (3) The designed chamber enabled the spray to operate at reduced pressure for increasing sprayed particle velocity. In the feasibility study, both suspensions of plasmid and plasmid-coated gold particles were used. The plasmid used was the commercially available Enhanced Green Fluorescent Protein gene. COS-1 cells were used as the target and the liquid media was evacuated immediately prior to the spraying process. Electrospraying was conducted at ambient pressure and the duration was no more than 2 min. After the spray transfection, the media was immediately replaced and the cell samples were returned to the incubator for 36 h. Transgene expression was detected by cellular fluorescence. This technology promises to have great potential for gene transfection and therapy studies.
Keywords:electrospray of nanodroplets  gene therapy  gene transfection  DNA transfer  gold nanoparticles  design of apparatus
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