Developing a Compartmentalized Co-Culture System to Create an In Vitro Model of Neuro-Immune Interaction

Abstract

The goal of this project was to develop a compartmentalized co-culture model that could be used to explore and manipulate the neuro-immune interaction that occurs during neurodegenerative diseases such as amyotrophic lateral sclerosis (ALS). While mouse and other animal models have given researchers a lot of insight into disease pathology, these models have major limitations. With the advent of the transformative stem cell technology, researchers have gained the capability to develop relevant human neuronal disease models. These disease models now form the basis of drug discovery screens. In order to assess the effects of neuro-immune interactions on neuronal projections versus cell bodies, which is relevant for ALS disease modeling, a compartmentalized system is required, preferably in a high throughput format. This study focuses on filling this gap by using stem cell technology in combination with an engineering approach to develop a compartmentalized in vitro system that allows for the investigation of neuro-immune interactions using human motor neurons. First motor neuron derived from human induced pluripotent stem cells (iPSCs) were plated directly with macrophages in a traditional co-culture setting which involves the two cell types having direct contact with each other. Next, the co-culture assay was developed in commercial Campenot chambers, which fluidically separated the two cell types while allowing the axons of the motor neurons to interact with the macrophages. Finally, several different designs, materials, and fabrication methods were explored to produce a novel compartmentalized chamber that would allow for the study of the neuro-immune interaction in a medium- to high-throughput manner.