The therapeutic potential of adipose tissue-derived mesenchymal stem cells (ADMSCs) is well studied for use in non-healing wounds. However, concerns on the transplantable cell number requirement, cell expansion, cell viability, retained cell multipotency and the limited cell implantation time for efficient impact hinders cell therapy. Recent literature is much inclined to the superiority of the ADMSCs' secretome, pre-dominating its paracrine-mediated therapeutic impact. In this context, the possibility of attaining accelerated wound angiogenesis through non-viral mediated enrichment of the ADMSCs secretome with pro-angiogenic growth factors (AGF) seems promising. Accordingly, this study aimed to explore the effect of AGF-enriched ADMSCs secretome for accelerating wound angiogenesis and repair in acute large area full thickness excision rabbit wound model, as adopted from Salgado et al. (Chir Buchar Rom 108:706-710, 1990). Using sub-dermal single-dose injections along the margin of the dorsal wound, native ADMSCs secretome, AGF-enriched ADMSC secretome, allogenic rabbit ADMSCs and a combination of AGF-enriched ADMSC secretome with allogenic rabbit ADMSCs were transplanted independently. Twenty-eight days (28 days) post-transplantation, histopathological analysis was performed to assess the effect. Hematoxylin and eosin (H&E) staining showed enhanced epithelization, notable granulation tissue and collagen fiber deposition in AGF-enriched secretome transplanted groups. This was confirmed by elevated CD31 detection, faster wound closure time and collagen organization. The use of single-dose AGF-enriched ADMSCs' secretome for therapeutic angiogenesis and wound repair seems to be a promising cell-free therapeutic option. Further investigations using multiple doses on larger animal groups remains to be explored in order to ascertain the comparative potential of AGF-enriched ADMSCs' secretome.