Chronic non-healing cutaneous wounds drastically reduce the quality of life for afflicted patients and costs healthcare systems over $50 billion annually. Clinical management of these wounds remains a challenge as current treatments are of limited efficacy. Recently, research in stem cell therapies for wound healing has started to gain more momentum. The aim of this study was to demonstrate that the stromal vascular fraction (SVF), which contains adipose-derived stem cells (ADSC), could accelerate keratinocyte wound closure without direct cell-cell contact and to identify potential cytokines, chemokines and growth factors involved. In-vitro wound closure assays were set up using primary human epidermal keratinocytes, primary (P0) SVF obtained from patients' fat grafts and their matching passaged (P3) SVF. SVF cells were plated on porous membranes of transwell inserts that prevent cell migration while allowing free diffusion of secreted proteins. Keratinocytes were placed in the bottom chamber with a scratch to simulate wound area. Scratch areas were monitored for 36h and following wound closure, media from each well was harvested and analyzed using a panel of 71 different cytokines, chemokines, and growth factors. Our results show that wound closure was accelerated 1.5-fold with P0-SVF and nearly 2-fold with P3-SVF compared to controls. Additionally, six cytokines were significantly upregulated in SVF-keratinocyte cultures during wound closure, namely; IL-6, IL-8, ENA-78, G-CSF, GRO-α, and MCP-1. Identification of secreted factors responsible for the acceleration of wound healing is the first step toward designing a standardized cell-free therapy to treat millions of patients with chronic non-healing wounds annually