A new method to fabricate porous chitosan nerve conduits with multi-channels was described. A uniquely designed mold was composed of 7-50 stainless steel needles and a set of plastic pedestals. Porous or imperforate chitosan tubes with 2-5mm inner diameter and 0.2-1.0 mm wall thickness were made firstly. The chitosan tubes were injected with 3% chitosan gel. The stainless steel needles longitudinally perforated through the chitosan tubes filled with chitosan gel, and the plastic pedestals were used to fix the needles. Lyophilization was used to finish fabrication. The diameter of channels was 0.2-0.4mm. Swelling property and biodegradability of Multi-channeled chitosan conduits were investigated. Wright’s staining and scanning electron microscope (SEM) were used to observe spread and proliferation of Neuroblastoma cells (N2A, mouse) on the conduits. It is promising that the porous chitosan nerve conduits with multi-channels are used as nerve tissue engineering scaffolds in repair of peripheral nerve and spinal cord injuries.