Fig. 8

Astrocyte-mediated patterning of neuronal networks. a Schematic illustration of the combination of micro-contact printing and microfluidics for patterning neurons and astrocytes together. b Immunocytochemistry showing that neuronal axons labeled by Smi312 (green) are guided by astrocytes GFAP (red). Astrocytes were patterned to extend long and slender processes in the horizontal direction. White arrows show where neuronal axons and astrocyte processes are adjacent to each other. c 3D confocal reconstruction of the neuronal networks formed by co-culturing neurons and astrocytes. Neurons are labeled with pan neuronal marker Tuj1, while astrocytes are labeled with GFAP. Black arrows show where neuronal processes and astrocyte processes are adjacent to each other. d The compass plot shows orientation of neurites from neurons adjacent to astrocyte processes. Most of the neurites go along the astrocyte processes (red 0°, n = 28) or along the channel edges (blue 90°, 270°, n = 17). e The length of neurites guided by the astrocytes is significantly longer than those growing along the channel (***P < 0.001, astrocyte: n = 28, channel: n = 17, Student’s t test). Scale bar 200 µm. Units in c is µm