Nanogroove-Enhanced Hydrogel Scaffolds for 3D Neuronal Cell Culture: An Easy Access Brain-on-Chip Model.

Bastiaens Bastiaens,Xie Xie,Luttge Luttge

doi:10.3390/mi10100638

Bastiaens Bastiaens, Xie Xie + Show 1 more

Open Access

https://doi.org/10.3390/mi10100638

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Abstract

In order to better understand the brain and brain diseases, in vitro human brain models need to include not only a chemically and physically relevant microenvironment, but also structural network complexity. This complexity reflects the hierarchical architecture in brain tissue. Here, a method has been developed that adds complexity to a 3D cell culture by means of nanogrooved substrates. SH-SY5Y cells were grown on these nanogrooved substrates and covered with Matrigel, a hydrogel. To quantitatively analyze network behavior in 2D neuronal cell cultures, we previously developed an automated image-based screening method. We first investigated if this method was applicable to 3D primary rat brain cortical (CTX) cell cultures. Since the method was successfully applied to these pilot data, a proof of principle in a reductionist human brain cell model was attempted, using the SH-SY5Y cell line. The results showed that these cells also create an aligned network in the 3D microenvironment by maintaining a certain degree of guidance by the nanogrooved topography in the z-direction. These results indicate that nanogrooves enhance the structural complexity of 3D neuronal cell cultures for both CTX and human SH-SY5Y cultures, providing a basis for further development of an easy access brain-on-chip model.

Highlights

Current models to study the brain and brain diseases are limited in their capabilities to translate findings toward the discovery of drugs that help treat these diseases [1]
The nanogrooved wells of the fabricated PDMS constructs for 3D neuronal cell cultures were measured to assess the nanogrooved pattern fidelity
We developed a method of adding structural network complexity in 3D in vitro neuronal cell models

Summary

Introduction

Current models to study the brain and brain diseases are limited in their capabilities to translate findings toward the discovery of drugs that help treat these diseases [1]. The so-called organ-on-chips (OOC) technology provides an opportunity to study human cells or organoids in a physiologically relevant microenvironment, potentially bridging the gap between current pre-clinical studies and human-based clinical trials [5,6]. To study the brain and brain diseases in an OOC platform, coined a brain-on-chip (BOC), we require a well-designed microsystem that can incorporate an environment for brain cells in a culture which mimics structural complexity in 3D [7]. While brain organoids can be cultured and exhibit such 3D structural complexity [11], there is little control over the location of regions, where specific brain cell types or structures can be generated or the actual control of the distal arrangement of cells to each other. The use of micro- and nanotechnology can aid the design of BOC platforms that offer more control of these parameters and potentially more reproducible experiments

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