Geometric micro-shapes facilitate trackless connections between human astrocytes

Abstract

Objective.Cell patterning approaches commonly employed to direct the cytoplasmic outgrowth from cell bodies have been via chemical cues or biomaterial tracks. However, complex network designs using these approaches create problems where multiple tracks lead to manifold obstructions in design. A less common but alternative cell patterning modality is to geometrically design the nodes to project the cytoplasmic processes into a specific direction, thus, removing the need for tracks. Janget alperformed an in-depth study of how rodent neuron primaries could be directed accurately using geometric micro-shapes. In parallel and in contrast, to the work of Janget alwe investigate, for the first time, the effect that micro-shape geometry has on the cytoplasmic process outgrowth of human cells of astrocyte origin using the biomaterial parylene-C.Approach.We investigated eight different types of parylene-C micro-shape on SiO₂substrates consisting of the: circle, square, pentagon, hexagon, equilateral triangle and three isosceles triangles with top vertex angles of 14.2°, 28.8°, and 97.6°, respectively. We quantified how each micro-shape influenced the: cell patterning, the directionality of the cytoplasmic process outgrowth and the functionality for human astrocyte.Main results.Human astrocytes became equally well patterned on all different micro-shapes. Human astrocytes could discriminate the underlying micro-shape geometry and preferentially extended processes from the vertices of equilateral triangles and isosceles triangles where the vertex angle equal to 28.8° in a repeatable manner whilst remaining functional.Significance.We demonstrate how human astrocytes are extremely effective at directing their cytoplasmic process outgrowth from the vertices of geometric micro-shapes, in particular the top vertex of triangular shapes. The significance of this work is that it demonstrates that geometric micro-shapes offer an alternative patterning modality to direct cytoplasmic process outgrowth for human astrocytes, which can serve to simplify complex network design, thus, removing the need for tracks.

Keywords: NT2; astrocytes; cell patterning; geometric micro-shapes; hNT; human; neural sprouting.