Microfluidic design for in vitro transfection of multicellular spheroids of HeLa cells using lipid-based gene delivery system

Abstract

Over the past decades, several comprehensive studies have led to the discovery of efficient treatment or prevention methods against genetic diseases. Among all methods, gene therapy, which is the introduction of suitable genetic material into specific cells to correct the abnormal genes, came out as a critical approach for this purpose. One of the crucial steps of gene therapy is understanding basic mechanism lying behind gene delivery or more specifically, the interaction between genetic material, vector system and target cells. However, in vivo studies are highly challenging due to several biological obstacles and in vitro 2D cell culture systems do not accurately represent dynamic in vivo models. In this context, 3D cell culture models, which mimic dynamic microenvironments of living organs, appear as a promising technique to study cell differentiation as well as cell response against controlled release systems in order to better understand molecular basis of diseases. Moreover, when comparing with conventional 3D cell culture systems, microfluidic approaches seem more advantageous since they allow high-throughput analysis with advanced real-time monitoring using microscopy techniques. Considering all these concepts, with this project, we aim to develop a cancer-on-a-chip based microfluidic platform that allows the transfection of cancer cells in spheroid form with small interfering RNA (siRNA) incorporated into lipid-based vector system. The transfection efficiency of the vector system as well as cell response will be monitored by fluorescent microscopy techniques and overall oxygen uptake rate of spheroids in microfluidic device will be determined by specific sensors. This project will be developed in collaboration with the Department of Biochemical Engineering of University College London in United Kingdom. The Microfluidic Research Group is led by Professor Nicolas Szita, who has broad expertise in advanced microfabrication techniques, Lab-on-a-chip systems, and system-wide integration of analytics. In this project, we expect to contribute in the field of lab-on-a-chip concept for dynamic cell study in 3D microenvironment.