A microfabricated multi-compartment device for neuron and Schwann cell differentiation

Eleonora De Vitis,Alessandro Polini,Angelo Quattrini,Lorenzo Moroni,Velia La Pesa,Giuseppe Gigli,Francesca Gervaso,Alessandro Romano

doi:10.1038/s41598-021-86300-4

Abstract

Understanding the complex communication between different cell populations and their interaction with the microenvironment in the central and peripheral nervous systems is fundamental in neuroscience research. The development of appropriate in vitro approaches and tools, able to selectively analyze and/or probe specific cells and cell portions (e.g., axons and cell bodies in neurons), driving their differentiation into specific cell phenotypes, has become therefore crucial in this direction. Here we report a multi-compartment microfluidic device where up to three different cell populations can be cultured in a fluidically independent circuit. The device allows cell migration across the compartments and their differentiation. We showed that an accurate choice of the device geometrical features and cell culture parameters allows to (1) maximize cell adhesion and proliferation of neuron-like human cells (SH-SY5Y cells), (2) control the inter-compartment cell migration of neuron and Schwann cells, (3) perform long-term cell culture studies in which both SH-SY5Y cells and primary rat Schwann cells can be differentiated towards specific phenotypes. These results can lead to a plethora of in vitro co-culture studies in the neuroscience research field, where tuning and investigating cell–cell and cell–microenvironment interactions are essential.

Highlights

Understanding the complex communication between different cell populations and their interaction with the microenvironment in the central and peripheral nervous systems is fundamental in neuroscience research
In order to investigate the basic functions of the nervous system and the pathogenic mechanisms of neurological disorders, it is of paramount importance to understand how molecular, physical and biochemical cues modulate the neuronal and non-neuronal cell dynamics[2]
The results showed that, among the different setup used, TS configuration leads to optimal seeding and cell attachment, with a homogeneous cell distribution and morphology in the channels and it was chosen to perform all the following test

Summary

Introduction

Understanding the complex communication between different cell populations and their interaction with the microenvironment in the central and peripheral nervous systems is fundamental in neuroscience research. We showed that an accurate choice of the device geometrical features and cell culture parameters allows to (1) maximize cell adhesion and proliferation of neuron-like human cells (SH-SY5Y cells), (2) control the inter-compartment cell migration of neuron and Schwann cells, (3) perform long-term cell culture studies in which both SH-SY5Y cells and primary rat Schwann cells can be differentiated towards specific phenotypes. These results can lead to a plethora of in vitro co-culture studies in the neuroscience research field, where tuning and investigating cell–cell and cell–microenvironment interactions are essential. Using a neuroblastoma cell line (SH-SY5Y) and primary Schwann cells, we tested several geometrical features and different microfluidic setups for controlling cell migration across the microchannels, evaluated the contribution of different adhesion molecules on cell seeding and proliferation, and performed cell differentiation on chip

Methods

Results

Conclusion