Advancing a MEMS-Based 3D Cell Culture System for in vitro Neuro-Electrophysiological Recordings

Bastiaens, Alex J.; Frimat, Jean-Philippe; Nunen, A.G.A. van; Schurink, Bart; Homburg, F.G.A.; Lüttge, Regina

doi:10.3389/fmech.2018.00021

Cited by 20 publications

(16 citation statements)

References 26 publications

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“…Conversely, the data shown in Table S1 suggests that the volume of Matrigel used for the 3D SH-SY5Y cell cultures was not detrimental with regard to nutrition supply, as the overall number of cells or outgrowths were within a similar range compared to 2D SH-SY5Y cell cultures. Nevertheless, an integrated microfluidic culturing chamber, additional to the 3D constructions, may provide improvement by creating a robust platform for the circulation of medium in the culture environment, and therefore may result in a higher consistency of the cell viability [44,45].…”

Section: Discussionmentioning

confidence: 99%

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

2019

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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.

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Section: Discussionmentioning

confidence: 99%

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

2019

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“…Scale bar denotes 1 cm. With permission from the open access article of Bastiaens et al (2018a , Figure 2C) under the CC-BY license.…”

Section: Resultsmentioning

confidence: 99%

“…In case of optical readout using fluorescent microscopy the microbioreactor can also be sealed against a simple flat glass plate, which we did for this study. Here, a variant of the original design of the microbioreactors was applied, which has been also published by us previously ( Bastiaens et al, 2018a ). In more detail, microbioreactors were fabricated using soft lithography of PDMS in poly(dimethyl methacrylate) (PMMA) milled molds and assembly by a surface mount technique.…”

Section: Methodsmentioning

confidence: 99%

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

2020

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Brain-on-chip (BOC) technology such as nanogrooves and microtunnel structures can advance in vitro neuronal models by providing a platform with better means to maintain, manipulate and analyze neuronal cell cultures. Specifically, nanogrooves have been shown to influence neuronal differentiation, notably the neurite length and neurite direction. Here, we have drawn new results from our experiments using both 2D and 3D neuronal cell culture implementing both flat and nanogrooved substrates. These are used to show a comparison between the number of cells and neurite length as a first indicator for valuable insights into baseline values and expectations that can be generated from these experiments toward design optimization and predictive value of the technology in our BOC toolbox. Also, as a new step toward neuronal cell models with multiple compartmentalized neuronal cell type regions, we fabricated microtunnel devices bonded to both flat and nanogrooved substrates to assess their compatibility with neuronal cell culture. Our results show that with the current experimental protocols using SH-SY5Y cells, we can expect 200 – 400 cells with a total neurite length of approximately 4,000–5,000 μm per 1 mm 2 within our BOC devices, with a lower total neurite length for 3D neuronal cell cultures on flat substrates only. There is a statistically significant difference in total neurite length between 2D cell culture on nanogrooved substrates versus 3D cell culture on flat substrates. As extension of our current BOC toolbox for which these indicative parameters would be used, the microtunnel devices show that culture of SH-SY5Y was feasible, though a limited number of neurites extended into microtunnels away from the cell bodies, regardless of using nanogrooved or flat substrates. This shows that the novel combination of microtunnel devices with nanogrooves can be implemented toward neuronal cell cultures, with future improvements to be performed to ensure neurites extend beyond the confines of the wells between the microtunnels. Overall, these results will aid toward creating more robust BOC platforms with improved predictive value. In turn, this can be used to better understand the brain and brain diseases.

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“…The microbioreactor consists of a microwell where biological elements (cells or bacteria) are usually placed and a surrounding channel that allows for flowing nutrients (i.e., oxygen) required for the proliferation of the biological elements. These two components are connected with a porous membrane (usually PDMS) to accommodate for the transfer of nutrients to the microwell [84]. The challenge arises when the biological elements located to points furthest from the nutrient flowing channel are not cultured as effectively as their neighboring elements closer to the channel are.…”

Section: Micromixing Methodsmentioning

confidence: 99%

Microfluidic Magnetic Mixing at Low Reynolds Numbers and in Stagnant Fluids

et al. 2019

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Microfluidic mixing becomes a necessity when thorough sample homogenization is required in small volumes of fluid, such as in lab-on-a-chip devices. For example, efficient mixing is extraordinarily challenging in capillary-filling microfluidic devices and in microchambers with stagnant fluids. To address this issue, specifically designed geometrical features can enhance the effect of diffusion and provide efficient mixing by inducing chaotic fluid flow. This scheme is known as “passive” mixing. In addition, when rapid and global mixing is essential, “active” mixing can be applied by exploiting an external source. In particular, magnetic mixing (where a magnetic field acts to stimulate mixing) shows great potential for high mixing efficiency. This method generally involves magnetic beads and external (or integrated) magnets for the creation of chaotic motion in the device. However, there is still plenty of room for exploiting the potential of magnetic beads for mixing applications. Therefore, this review article focuses on the advantages of magnetic bead mixing along with recommendations on improving mixing in low Reynolds number flows (Re ≤ 1) and in stagnant fluids.

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Advancing a MEMS-Based 3D Cell Culture System for in vitro Neuro-Electrophysiological Recordings

Cited by 20 publications

References 26 publications

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

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

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

Microfluidic Magnetic Mixing at Low Reynolds Numbers and in Stagnant Fluids

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