This study aimed to investigate the role of a low dose of human umbilical cord blood mesenchymal stem cells (HUCB-MSCs) with chitosan scaffold (ChSc) on the progress of the cutaneous wound healing process in male Albino rats. A full-thickness cutaneous wound with a circular shape with a diameter of about 2 cm was induced on the dorsum of thirty-six Albino male rats, divided into four groups: the first group, the wounds were left without treatment as a control group. In the second group, the wounds were covered by freeze-dried ChSc. While in the third group, the wounds were treated through injection of the HUCB-MSCs intradermally, and in the fourth group, the ChSc seeded with HUCB-MSCs were used together to treat the wounds. The progress of wound healing was monitored by histological, immunohistochemical, and biochemical assays for all experimental groups at 3, 8, and 21 postoperative days. Both ChSc and the low dose of HUCB-MSCs alone performed moderate healing progress. The (HUCB-MSCs) ChSc group exhibited an increased healing rate more than the other groups and reported an appropriate collagen deposition without scarring signs, effective mast cell regulation, and well vascularization. In conclusion, the fourth group (the HUCB-MSCs with ChSc) improved the healing process, revealing the highest healing rate and performance without complications.