To prepare microplates for cellular seeding, the majority of the distilled water in all wells was removed, with a small amount (~10 μl per compartment) remaining in each well to preserve hydrophilicity. Microplates were angled toward the UV bulb and sterilized under UV radiation for 1 hour. Afterward, Matrigel™ (Corning) was added to cover the cell culture surface in each well. The devices were placed in the incubator for approximately one hour. Glia isolated from P0 pups were seeded into the wells one day prior (day 4–5) to iN seeding (day 5–6) with a glial cell concentration of approximately 300 cells per mm2. Glial cells were removed from the flask using Trypsin/EDTA and re-seeded into the microdevice in DMEM with 10% FBS and 1% P/S. In the microplate, remaining water was removed from the well and a 15 μl bolus of cells was added to the bottom of each compartment (3200 cells per 15 μl). Approximately one hour was given for the cells to adhere to the surface, after which approximately 200 μl of media was added to the well. This additional step was necessary to ensure cell restriction in their respective compartments.
