A combinatorial approach was used to present primary neurons with a large library of topographical features in the form of micropatterned substrate for high-throughput screening of physical neural-guidance cues that can effectively promote different aspects of neuronal development, including axon and dendritic outgrowth. Notably, the neuronal-guidance capability of specific features was automatically identified using a customized image processing software, thus significantly increasing the screening throughput with minimal subjective bias. Our results indicate that the anisotropic topographies promote axonal and in some cases dendritic extension relative to the isotropic topographies, while dendritic branching showed preference to plain substrates over the microscale features. The results from this work can be readily applied towards engineering novel biomaterials with precise surface topography that can serve as guidance conduits for neuro-regenerative applications. This novel topographical screening strategy combined with the automated processing capability can also be used for high-throughput screening of chemical or genetic regulatory factors in primary neurons.
In this study, we explored the concept of introducing asymmetry to cell shapes by patterned cell culture substrates, and investigated the consequence of this induced asymmetry to cell migration behaviors. Three patterns, named "Squares", "Grating", and "Arcs" were fabricated, representing different levels of rotational asymmetry. Using time-lapse imaging, we systematically compared the motility and directionality of mouse osteoblastic cells MC3T3-E1 cultured on these patterns. Cells were found to move progressively faster on "Arcs" than on "Grating", and cells on "Squares" were the slowest, suggesting that cell motility correlates with the level of rotational asymmetry of the repeating units of the pattern. Among these three patterns, on the "Arcs" pattern, the least symmetrical one, cells not only moved with the highest velocity but also the strongest directional persistence. Although this enhanced motility was not associated with the detected number of focal adhesion sites in the cells, the pattern asymmetry was reflected in the asymmetrical cell spreading. Cells on the "Arcs" pattern consistently displayed larger cytoplasmic protrusion on one side of the cell. This asymmetry in cell shape determined the direction and speed of cell migration. These observations suggest that topographical patterns that enhance the imbalance between the leading and trailing fronts of adherent cells will increase cell speed and control movement directions. Our discovery shows that complex cell behaviors such as the direction of cell movement are influenced by simple geometrical principles, which can be utilized as the design foundation for platforms that guide and sort cultured cells.
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