Compartmentalized neuronal cultures.

The culturing of neurons results in formation of the layer of neurons with random extensive overlapping outgrowth. To understand specific roles of somas, axons, and dendrites in complex function of neurons and to identify molecular mechanisms of biological processes in these cellular compartments various methods were developed. We utilized AXon Investigation System (AXIS™) manufactured by Millipore. This device provides an opportunity to orient neuronal outgrowth and spatially isolate neuronal processes from neuronal bodies. AXIS device is a slide-mounted microfluidic system, which consists of four wells. Two of the wells are connected by a channel on each side of the device. Channels are connected by microgrooves (approximately 120). The size of microgrooves (10 μm in width and 5 μm in height) do not permit passage of cell through while allowing extension of neurites. The microfluidic design also allows an establishment of a hydrostatic gradient when the volume in one chamber is greater than that in the other (Park et al. Nat Protocols 1: 2128-2136, 2006). This feature allows studying of the effect of specific compounds on selected compartments of neurons.