Spinal cord decellularized scaffolds can promote axonal regeneration and restore hindlimb motor function of spinal cord defect rats. However, Scarring caused by damage to the astrocytes at the margin of injury can hinder axon regeneration. Olfactory ensheathing cells (OECs) integrate and migrate with astrocytes at the site of spinal cord injury, providing a bridge for axons to penetrate the scars and grow into lesion cores. The purpose of this study was to evaluate whether decellularized scaffolds carrying OECs could better promote axon growth. For these studies, decellularized (DC) scaffolds were co-cultured with primary extracted and purified OECs. Immunofluorescence and electron microscopy were used for verification of cells adhere and growth on the scaffold. Scaffolds with OECs were transplanted into rat spinal cord defects to evaluate axon regeneration and functional recovery of hind limbs. BBB scoring was used to assess motor function recovery, and GFAP and NF200 stained tissue sections were used to evaluate axonal regeneration and astrological scar distribution. Our results indicated that spinal cord decellularized scaffolds have good histocompatibility and spatial structure, and can promote the proliferation of adherent olfactory ensheathing cells. In animal experiments, scaffolds carrying OECs have better axon regeneration promoting protein expression than the SCI model, and improve the proliferation and distribution of astrocytes at the site of injury. These results proved that the spinal cord decellularized scaffold with OECs can promote axon regeneration at the site of injury, providing a new basis for clinical application.This article is protected by copyright. All rights reserved.