The in vitro models developed to investigate the growth and myelination of axons, such as dorsal root ganglion (DRG)-Schwann cell co-culture, DRG-oligodendrocyte co-culture and central nervous system (CNS) neuron-oligodendrocyte co-culture, have provided an effective way to reveal the mechanisms that underlie the interaction between neurons and myelin-forming cells. In order to better understand the complex process of myelination during CNS development and spinal cord repair, we established a rat spinal cord neuron-oligodendrocyte co-culture model. In this co-culture system, the spinal cord explants were used as the source of neurons, and the oligodendrocytes were induced from GFP-oligodendrocyte precursor cells (GFP-OPCs). The results showed that the GFP-oligodendrocytes that differentiated from GFP-OPCs in co-culture attached to the neurites growing out from the spinal cord explants and formed myelin structures. As the oligodendrocytes expressed GFP, and the neuron somas remained in the explants, the interaction between oligodendrocytes and neurites in co-culture were observed clearly and dynamically without immunostaining.