Abstract

3D Organoids-Based High Content Screening

Xavier Gidrol, Laboratory Director, CEA Grenoble

To analyze the phenotypic consequences of genetic perturbation in mammalian cells with drugs, RNAi or expression vectors, there are increasing needs for systematic cell-based high content screening (HCS) approaches. Although several groups are performing HCS in human cell lines, the real challenge in translational research remains screening on a reduced number of primary cells directly obtained from patients in a microenvironment that would resemble the original tissues. While standard monolayer two-dimensional culture conditions are poor mimics of the cellular environment in-situ, microfabricated systems enable three-dimensional organoids cultures and have the potential to provide biological insight not achievable before. We use microfluidics and MEMS (MicroElectroMechanical Systems) to analyze the phenotypic consequences of genetic perturbations (RNAi-based HCS) in 3D organoids. Specifically, we present a novel approach based on a flow-focusing microfluidic system that encapsulates either single prostatic or mammary cell in Matrigel beads and assay for development of organoids. Furthermore, we developed new imaging technology to monitor live organoids self-assembly and inter organoids cell trafficking.