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

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.

Nanogrooves for 2D and 3D Microenvironments of SH-SY5Y Cultures in Brain-on-Chip Technology

“…With our image-based screening method we found that certain pattern dimensions increase organization within the cell culture through the alignment of neurites on the underlying nanogrooved substrate, positively correlating to neuronal differentiation and outgrowth length ( Bastiaens et al, 2018b , 2019a ). Also, the effect of the nanogrooves is extended into the 3D microenvironment as seen for primary rat cortical cells and SH-SY5Y cells on a nanogrooved substrate and within a hydrogel, here growth factor reduced Matrigel ( Bastiaens et al, 2019b ). As a first step in the optimization procedure for the BOC system design and its robustness, the effect of implementing features such as nanogrooves was quantified for total neurite length in 2D and 3D neuronal cell culture on nanogrooved substrates as a baseline expectation.…”

Astrocyte‐Encapsulated Hydrogel Microfibers Enhance Neuronal Circuit Generation

“…Albeit great clinical potential, glial transplantation still has several problems, including cell type and maturity of astrocytes, and, more severely, astrocyte loss after transplantation . A few reports have been found on the physically isolated confinement of astrocytes for 3D neuron‐culture platform, composed of various hydrogels including collagen, Matrigel as well as the synthetic peptides, to support and promote the neuronal development and neuronal circuit generation, while protecting astrocytes from the outside and preventing cell leakage and loss . However, the hydrogel scaffolds, used in the previous works, are macro‐sized (usually millimeter‐ or centimeter‐scaled).…”

“…[17] Albeit great clinical potential, glial transplantation still has several problems, including cell type and maturity of astrocytes, and, more severely, astrocyte loss after transplantation. [18] A few reports have been found on the physically isolated confinement of astrocytes for 3D neuron-culture platform, composed of various hydrogels including collagen, [19] Matrigel [20] as well as the synthetic peptides, [21] to support and promote the neuronal development and neuronal circuit generation, while protecting Astrocytes, the most representative glial cells in the brain, play a multitude of crucial functions for proper neuronal development and synaptic-network formation, including neuroprotection as well as physical and chemical support. However, little attention has been paid, in the neuroregenerative medicine and related fields, to the cytoprotective incorporation of astrocytes into neuron-culture scaffolds and full-fledged functional utilization of encapsulated astrocytes for controlled neuronal development.…”

Editorial for the Special Issue on Organs-on-Chips

“…Since the cost of drug discovery is constantly increasing due to the limited predictability of conventional monolayer culture methods and animal models, this technology has great potential to promote drug discovery and development as well as to model human physiology and disease.This Special Issue is themed to provide insight and advancements in organ-on-chip microdevices. There are fifteen papers including three review papers, covering a novel material to fabricate microfluidic organs-on-chips [1], methods to deliver mechanical stimuli [2,3], methods to measure mechanical forces [4,5], methods to evaluate cellular functions in 3D cultures [6][7][8], and specific organ models; lung chips [3,9], liver chips [10,11], blood vessel chips [12][13][14][15] including models of the outer blood-retina barrier [14] and ischemia-reperfusion injury [15].Inside the body, cells are exposed to biomechanical forces, including fluidic shear stress and mechanical strain, which regulate cell function and contribute to disease. Kaarj et al reviewed methods to produce mechanical stimuli focusing on the technical details of devices [2].…”

“…By combining 3D cell culture and microdevice technologies, 3D culture devices have been developed to replicate in vivo-like microenvironment as well as to develop high-throughput systems. Bastiaens et al developed a 3D neuronal culture device by combining nanogrooved substrates with a 3D hydrogel culture [6]. This method permits the formation of an aligned 3D neural cell network.…”