Microfluidic droplets device to transfect non-adherent mammalian cells in vitro

• Grant number: 14/10557-5
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate
• Start date: September 01, 2014
• End date: August 31, 2015
• Field of knowledge: Interdisciplinary Subjects
• Principal Investigator: Lucimara Gaziola de la Torre
• Grantee: Micaela Tamara Vitor
• Supervisor: Charles N. Baroud
• Host Institution: Faculdade de Engenharia Química (FEQ). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil
• Institution abroad: École Polytechnique, France
• Associated to the scholarship: 12/24797-2 - Microfluidic droplets devices to incorporate nucleic acids into cationic liposomes and chitosan nanoparticles and to transfect mammalian cells in vitro, BP.DR

Abstract

This project aims to develop a microfluidic droplets system to transfect non-adherent mammalian cells, more specifically Jurkat T cells, using nanoparticles as nucleic acid nanocarriers. For this, nucleic acids encoding the green fluorescent protein will be incorporated into cationic liposomes and chitosan nanoparticles, and then inserted in the microfluidic droplets system with non-adherent mammalian cells, to induce the transfection within the droplets. Traditional case of stagnant flow transfection in wells allows only diffusive transport of nanoparticles/nucleic acid to the cell surface; in contrast, the convective contribution in micro-droplets facilitates and enhances the control of the transfection. Additionally, droplet-based microfluidics demonstrated as a potentially more sensitive method for biomarker discovery than conventional microfluidic systems, since the encapsulation permits amplified detection of extremely low levels of biomarker molecules. Furthermore, based on the "rails and anchors" channel design, the motion of droplets can be control, optimizing the number of cells that can be observed and the flow within the stationary droplets to control the shear stress that is felt by the cells while they remain in the field of view of the microscope. Thus, this system will enable us to study the optimum molar ratio between the nanoparticles and nucleic acids, the rate between nanoparticle/cell and the effect of shear stress inside the droplets on transfection. Then, the transfection occurred in the microfluidic system will be compared to the traditional transfection, performed in wells (bulk method). This project will be developed by networking our research group that is expertise in nanoparticles production and the international collaboration of Dr. Charles N. Baroud from École Polytechnique in France. He coordinates a research group that developed several lines of applications to lab-on-a-chip devices, including the cell biology in droplets. This project will contribute in nanobiotechnology, microfluidics and gene delivery areas. (AU)

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

VITOR, MICAELA TAMARA; SART, SEBASTIEN; BARIZIEN, ANTOINE; DE LA TORRE, LUCIMARA GAZIOLA; BAROUD, CHARLES N.. Tracking the Evolution of Transiently Transfected Individual Cells in a Microfluidic Platform. SCIENTIFIC REPORTS, v. 8, JAN 19 2018. (14/10557-5, 15/26701-0)