Nerve guidance conduits (NGCs) are an essential solution for peripheral nerve repair and regeneration in tissue engineering and medicine. However, the ability of current NGCs is limited to repairing longer nerve gap (i.e., >20 mm) because it cannot meet the following two conditions simultaneously: (1) directional guidance of the axial high-density channels and (2) regenerative stimulation of the extracellular matrix secreted by Schwann cells (SCs). Therefore, we propose a multi-material 3D bioprinting process to fabricate multi-channel nerve guide conduits (MNGCs) containing SCs. In the article, cell-laden methacrylate gelatin (GelMA) was used as the bulk material of MNGCs. To improve the printing accuracy of the axial channels and the survival rate of SCs, we systematically optimized the printing temperature parameter based on hydrogel printability analysis. The multi-material bioprinting technology was used to realize the alternate printing of supporting gelatin and cell-laden GelMA. Then, the high-accuracy channels were fabricated through the UV cross-linking of GelMA and the dissolving technique of gelatin. The SCs distributed around the channels with a high survival rate, and the cell survival rate maintained above 90%. In general, the study on multi-material 3D printing was carried out from the fabricating technology and material analysis, which will provide a potential solution for the fabrication of MNGCs containing SCs.