Abstract
Conventional in vitro cultures are useful to represent simplistic neuronal behavior; however, the lack of organization results in random neurite spreading. To overcome this problem, control over the directionality of SH-SY5Y cells was attained, utilizing photolithography to pattern the cell-repulsive anionic brush poly(potassium 3-sulfopropyl methacrylate) (PKSPMA) into tracks of 20, 40, 80, and 100 μm width. These data validate the use of PKSPMA brush coatings for a long-term culture of the SH-SY5Y cells, as well as providing a methodology by which the precise deposition of PKSPMA can be utilized to achieve a targeted control over the SH-SY5Y cells. Specifically, the PKSPMA brush patterns prevented cell attachment, allowing the SH-SY5Y cells to grow only on noncoated glass (gaps of 20, 50, 75, and 100 μm width) at different cell densities (5000, 10 000, and 15 000 cells/cm2). This research demonstrates the importance of achieving cell directionality in vitro, while these simplistic models could provide new platforms to study complex neuron–neuron interactions.
Highlights
Organized architectures with defined pathways are known to be present in the nervous system, for example, chick and mouse dorsal retina comprise defined canals, which are packed with axons.[1,2] in frogs, the dorsal column provides tracks that guide the dorsal root ganglion axons after their entry into the spinal cord.[3]
A change of color on the coated silicon wafers when compared to the noncoated silicon wafers resulted in the optical observation of the channels, where a gray/blue pattern could be observed when the photolithography process was carried out (Figure S3)
The medium cell density (10 000 cells/cm2) seemed more consistent among the different parameters used, resulting in both cell−cell contact and the possibility of neurite spreading. These results indicated that cell density did not affect significantly the different channel sizes and that the 20 μm channel width showed the constraint of elongated cells and was suitable for obtaining neurite directionality
Summary
Organized architectures with defined pathways are known to be present in the nervous system, for example, chick and mouse dorsal retina comprise defined canals, which are packed with axons.[1,2] in frogs, the dorsal column provides tracks that guide the dorsal root ganglion axons after their entry into the spinal cord.[3] neuronal directionality is present during development, but it is essential in neural regeneration. Studies have demonstrated that neurons are highly influenced by their surroundings, indicating a strong interaction at the interface between the cell and the material surface[6−8] and a high sensitivity to the changes in their external environment. Rajnicek et al used primary Xenopus spinal cord and rat hippocampal neurons to investigate the variations in neuronal guidance through parallel grooves of various widths (1, 2, and 4 μm) and depths (14− 1100 nm) produced by electron beam lithography.[1]
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