Schwann cells(SCs), an important component of the peripheral nervous system, interact with neurons to mutually support growth and replication for the peripheral nerve regeneration. Recently, adding SCs to the lumen of guidance channel is widely tried to improve regeneration or to make regeneration possible over otherwise irreparable gaps. However, it is not easy to isolate and multiplicate SCs as much as enough to help the axonal regeneration. For the allogeneic SCs source for tubular nerve guidance, we developed a little bit improved technique of harvesting and multiplicating SCs. By culturing dispersed dorsal root ganglia in specially designed medium with growth factors and serial processing, we repeatedly generate relatively homogenous SC cultures. Our technique was compared with other methods of literature using immunostaining methods such as GFAP, S100, BDNF and the total SC count assessment at different time interval after primary culture.