AUTHOR=Bastiaens Alex , Sabahi-Kaviani Rahman , Luttge Regina TITLE=Nanogrooves for 2D and 3D Microenvironments of SH-SY5Y Cultures in Brain-on-Chip Technology JOURNAL=Frontiers in Neuroscience VOLUME=Volume 14 - 2020 YEAR=2020 URL=https://www.frontiersin.org/journals/neuroscience/articles/10.3389/fnins.2020.00666 DOI=10.3389/fnins.2020.00666 ISSN=1662-453X ABSTRACT=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 substrate. 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 towards design optimization and predictive value of the technology in our BOC toolbox. Also, as a new step in towards 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 4000 – 5000 µm per 1 mm2 within our BOC devices, with the exception of 3D neuronal cell cultures on flat substrates. Whilst culture of SH-SY5Y was feasible within the microtunnel devices, only limited numbers of neurites extended into microtunnels away from the cell bodies, regardless of using nanogrooved or flat substrates. 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.