A simple, yet versatile, co-culture method for examining neuron-glia interactions

Abstract There is increasing interest in examining neuron-glial interactions in the developing and the mature central nervous system. Here, we describe a system that allows examination of interactions mediated by diffusible molecules even when such interactions involve more than 2 cell populations. Our procedure involves culturing highly enriched populations of neurons and glial cells on separate coverslips. Such coverslips can be readily moved from one petri dish to another, thus enabling the investigator to subject the neurons and the glial cells to different experimental manipulations. The coverslips can then be placed in any desired combination within a petri dish. This gives one great flexibility in examining neuron-glial interactions. Using this approach, we unequivocally demonstrate that astrocytes protect neurons from glutamate excitotoxicity.