Peripheral nerves conducting motor and somatosensory signals in vertebrate consist of myelinated and unmyelinated axons.In vitromyelination culture, generated by co-culturing Schwann cells (SCs) and dorsal root ganglion (DRG) neurons, is an indispensable tool for modeling physiological and pathological conditions of the peripheral nervous system (PNS). This technique allows researchers to overexpress or downregulate molecules investigated in neurons or SCs to evaluate the effect of such molecules on myelination.In vitromyelination experiments are usually time-consuming and labor-intensive to perform. Here we report an optimized protocol forin vitromyelination using DRG explant culture. We found that ourin vitromyelination using DRG explant (IVMDE) culture not only achieves myelination with higher efficiency than conventionalin vitromyelination methods, but also can be used to observe Remak bundle and non-myelinating SCs, which were unrecognizable in conventional methods. Because of these characteristics, IVMDE may be useful in modeling PNS diseases, including Charcot Marie Tooth disease (CMT),in vitro. These results suggest that IVMDE may achieve a condition more similar to peripheral nerve myelination observed during physiological development.