Method for analysis of electrospray for gene transfer and the impact on cell viability of A549 alveolar epithelial like cells

Electrospray is a process based on creation and acceleration of small sized droplets based on electrostatic repulsion. Spraying plasmid containing liquids this process may be used to transfer genes into cells. Within this paper we report on a method for accessing and evaluating the spray modalities using high speed imaging system with a post processing of image data to obtain estimated volume and velocity of emerging droplets first. Second we investigate on the impact of different media on the spray modalities. Third we evaluate the impact of the spray on cell viability and on transfection efficiency of an eGFP plasmid as reporter gene obtained in an in vitro setup on alveolar epithelial like cells (A549).

[1]  Xiang Gao,et al.  Nonviral Gene Delivery: Principle, Limitations, and Recent Progress , 2009, The AAPS Journal.

[2]  S. Gaskell Electrospray: Principles and Practice , 1997 .

[3]  Hong Shen,et al.  The morphometrical analysis on the ultrastructure of A549 cells. , 2010, Romanian journal of morphology and embryology = Revue roumaine de morphologie et embryologie.

[4]  Takafumi Sakai,et al.  DNA introduction into living cells by water droplet impact with an electrospray process. , 2008, Angewandte Chemie.

[5]  M. Detamore,et al.  Physical Non-Viral Gene Delivery Methods for Tissue Engineering , 2012, Annals of Biomedical Engineering.

[6]  James M. Wilson,et al.  Methods of gene delivery. , 1998, Hematology/oncology clinics of North America.

[7]  S. Šatkauskas,et al.  A new electrospray method for targeted gene delivery , 2018, Scientific Reports.

[8]  J. MacQueen Some methods for classification and analysis of multivariate observations , 1967 .

[9]  David Hradetzky,et al.  An approach towards bronchoscopic-based gene therapy using electrical field accelerated plasmid droplets , 2012, 2012 Annual International Conference of the IEEE Engineering in Medicine and Biology Society.