31966–021), 10% FCS, NEAA (100x, Sigma, M7145), Pen/strep (100x)) for about 5–6 days. At this point (day 7 of the astrocytes), neurons were plated directly on astrocyte coverslips for direct contact mode. For indirect contact mode, neurons were plated on PLO/ laminin-coated coverslips and the next day (day 19) flipped over with neuron side (down) towards astrocytes in the indirect contact wells. For indirect contact cultures, the neuron coverslips were replaced into plates with freshly plated astrocytes after 3 weeks of co-culturing. Animals were housed and bred according to institutional, Dutch and U.S. governmental guidelines. The institutional ethical committee of the VU University approved these studies.
