Microfluidic droplets devices to incorporate nucleic acids into cationic liposomes and chitosan nanoparticles and to transfect mammalian cells in vitro

Abstract

This project aims to develop microfluidic droplets systems to incorporate nucleic acids encoding green fluorescent protein in cationic liposomes and chitosan nanoparticles, and also to use these same systems to transfect mammalian cells, T lymphocytes and dendritic cells, through these nanostructures. For this, the first step will be the development of a microfluidic droplets system to incorporate nucleic acids into the nanoparticles, since microfluidic systems based on droplets promote rapid mixing, generating complexes with smaller size and polydispersity. In the second step, microfluidic systems will be developed to transfect mammalian cells within droplets. Traditional case of stagnant flow transfection in wells allows only diffusive transport of nanoparticles, in contrast, the convective contribution in microfluidic systems facilitates and enhances the control of transfection. Additionally, in the microfluidic droplet system, there is a larger interfacial area per unit volume, allowing a faster mass transfer than in a laminar flow-based system. Thus, these microfluidic droplets systems will enable us to study the optimum molar ratio between the nanoparticles and nucleic acids, the distribution of nanoparticles per cell and the shear stress inside the droplets for transfection. Then, the transfection occurred in the microfluidic droplets system will be compared to the traditional transfection performed in wells. This project will be developed by our research group, expertise in nanoparticles production, assisted in biological area by Dr. Ronei Luciano Mamoni, a researcher at the Faculty of Medical Sciences of Unicamp, and in collaboration with Prof. Dr. Charles N. Baroud research group from École Polytechnique in France, expertise in microfluidic droplets system for biological applications. This project will contribute in nanobiotechnology, microfluidics and gene delivery areas. (AU)