Colloids as Mobile Substrates for the Implantation and Integration of Differentiated Neurons into the Mammalian Brain
Figure 1
Development and manipulation of neurons supported on silica beads.
Confocal microscopy z series are projected on the xy scanning plane. E18 hippocampal neurons cultured seeded at 4k cells/cm2 on 125 µm (a-c), and on 45 µm (d-f) PLL coated beads shown at DIV 4. Cells were fixed and stained with a neuron specific alphãtubulin antibody (green), and with an axon specific smi-312 antibody (red). Neurons were polarized in both preparations independently of bead radius of curvature. The number of neurons per bead is proportional to bead surface area, as 45 µm beads carried on average one cell, and 125 µm beads carried about 10 cells. (g) Bright field image of neurons seeded at 100k cells/cm2 on 45 µm beads at DIV 4. (h) Cells were fixed and stained with a neuron specific Tuj-1 antibody (red), and the nuclear marker DAPI (blue). Twenty-one of the twenty-five cells on this bead are Tuj-1 positive. At this high density, cells in direct contact with the bead surface wrap their processes around the beads (highlighted in red) while the others sit on this layer (highlighted in blue) as illustrated in the color-coded picture (i). All Scale bars = 50 µm.