A Co-culture Method to Model Neuron-Oligodendrocyte Interactions

On day 14, plate iOPCs at a density of 1 x 10⁵ cells per well in a 24-well plate in OPC differentiation medium. On the next day, detach the induced human neurons with cell detachment solution. Add neurons onto the cultured OPCs, plating at the seating density of 2 x 10⁵ cells per well. Use co-culture medium containing neurobasal A-medium, 2% B-27 Supplement, and 100 nanograms per milliliter T3 triiodothyronine. Change the medium on the next day, and then every other day. If OPCs proliferate too fast and reach confluency in less than three days, add 2 to 5 micromolar Ara-C.