Diabetes is associated with numerous complications, such as diabetic skin wounds or ulcerations. The aim of this study was to evaluate experimentally the effectiveness of applying polycaprolactone (PCL)‐gelatin scaffold, with or without rat CD93 hematopoietic stem cells (HSCs), in diabetic wound healing in a rat model. CD93 HSCs were aseptically isolated from rat bone marrow using fluorescent activated cell sorting (FACS) method and FACS‐SORTER. A total of 25 Wistar rats were divided into five groups including Group I (sham, nondiabetic, and wound covered only with sterile dressing), II (control, diabetic rat), III (CD93 HSCs alone), IV (PCL‐gelatin scaffold), and V (CD93 HSCs+PCL‐gelatin scaffold). Animals were killed on Days 7, 14, or 28 posttreatment and histological sections were blindly evaluated by two expert pathologists. Death‐associated protein kinase 1 (DAPK‐1) gene and vesicular endothelial growth factors (VEGF) protein expression were evaluated using reverse transcription‐polymerase chain reaction and western blot, respectively. The thickest and the thinnest epidermises microscopically were belonged to CD93+HSCs+scaffold and the control group, respectively. The growth rate of the epidermis and adnexal epithelia was the highest in both the cell and cell+scaffold groups. Evaluation of the protein expression level of VEGF indicated that the expression levels of this growth factor were the most on Day 7 posttreatment in sham, HSCs alone, and HSCs cell+scaffold groups. While the lowest expression levels of this growth factor was detected in the control and scaffold groups. The gene expression level of DAPK‐1 on Day 7 posttreatment was higher than that of the Day 14 posttreatment in all groups. The highest and lowest gene expression levels of DAPK‐1 belonged to control and sham groups, respectively. According to our findings, CD93 HSCs offer new prospects for the treatment of diabetic ulcers and concomitant application of these cells with PCL‐gelatin nanofiber scaffold significantly improves diabetic wound treatment. Based on the findings of the present study, it can be concluded that the polycaprolactone (PCL)‐gelatin scaffold associated with CD93+HSCs could act as an appropriate tissue engineering construct in diabetic wound healing. The CD93 HSCs present adhesion, growth, and proliferation properties in the vicinity of the PCL‐gelatin nanofiber scaffold. Despite delayed wound healing in diabetic ulcer, this compound can accelerate the wound healing process of diabetic wounds in rat. This property is due to the stimulation of vesicular endothelial growth factors secretion at the wound site as well as stimulation of angiogenesis for early regeneration, higher collagenization rate, epidermis formation, and thickening of the dermis in less time. Moreover, it is associated with the further growth of hair follicles at the site of the healed wound. Meanwhile, the results of this study confirmed the presence of CD93 HSCs at the wound site, even after 28 days of transplantation. This study also demonstrated that death‐associated protein kinase 1 gene plays an inhibitory role in the diabetic wound healing process.