Simultaneous neurite elicitation and elongation from neurons using a microfabricated post array

We report the use of a microfabricated post array with supporting pillars to apply direct mechanical force to initiate and elongate multiple neurites from chick forebrain neurons in vitro. The micropost array was placed on top of the cell culture on a glass cover slip; a gap of 3-5μm was achieved between the tips of the posts and the cell culture surface substrate due to the differential height of the posts and the supporting pillars. After allowing ∼1.5 hours for the cells to adhere to the posts, the post array was moved laterally with respect to the substrate and cells at a constant velocity of 36μm/hr. Several neurite-like cytoplasmic processes initiated from cells that had formed adhesion to the posts, and these neurites elongated in response to further translation. This is the first report of the use of a microfabricated device to simultaneously apply mechanical force to multiple neurons to elicit and elongate neurites. We envision that this technique will enable the construction of living neural networks having defined connectivity.