Intraoperative Rapid Harvest of Autologous Epidermal Stem Cells Improves Single-Stage Composite Skin Graft Survival in Full-Thickness Wounds

BACKGROUND: Full-thickness wounds, especially in functional areas, require grafts with high survival potential and minimal contraction risk. Composite skin grafts combining split-thickness skin grafts (STSGs) with acellular dermal matrix (ADM) offer improved outcomes but are limited by delayed vascularization, requiring staged procedures. The authors developed tissue-engineered skin (TES) using intraoperative Myseed technology-isolated epidermal stem cells (ESCs) to enhance ADM vascularization and graft survival. METHODS: ESCs were isolated from rat STSGs using Myseed technology and assessed using flow cytometry and CD44/CD90/CD106 markers. Full-thickness dorsal wounds (2.5 cm) in 24 Sprague Dawley rats were randomized into a TES group (ADM + ESCs + STSGs), a composite skin (CS) grafts group (ADM + STSGs), and an STSG group. Outcomes included survival, contraction, angiogenesis (evaluated using laser speckle imaging and CD31-positive vessel density), and histologic results at days 4, 7, 14, and 21. RESULTS: The concentration of ESCs extracted from STSGs was approximately 1.03 × 105/cm2, and cell activity was approximately 83.7%. The TES group showed superior survival, with a failure rate of 1 of 8 versus 3 of 8 in the CS group, achieving full closure by day 21 with a nearly normal texture. The TES group had reduced contraction (20.85% versus 39.48% in the STSG group; P < 0.001) and accelerated healing on days 7, 14, and 21 (P < 0.001). Angiogenesis was enhanced in the TES group, with higher perfusion on days 4, 7, and 14 (P < 0.01) and a greater number of CD31-positive vessels on day 7 than in the CS group (P < 0.01). CONCLUSION: Myseed technology facilitates single-stage TES construction by rapidly isolating ESCs, addressing ADM vascularization delays, improving graft survival, and reducing contraction, offering an efficient solution for full-thickness wound repair, especially in functional areas. CLINICAL RELEVANCE STATEMENT: This study provides the first evidence that intraoperative isolation of ESCs using Myseed technology facilitates rapid TES construction, enhancing ADM vascularization and graft outcomes. It offers a practical, single-stage alternative to traditional methods for treating full-thickness functional skin defects.