Keratinocytes and Adipose-Derived Mesenchymal Stem Cells: The Heir and the Spare to Regenerative Cellular Therapies for Difficult-to-Heal Skin Wounds

This dissertation by Hady Shahin explores the development of a bio-production workflow for autologous keratinocyte-based Advanced Therapy Medicinal Products (ATMPs) and investigates the potential of adipose-derived mesenchymal stem cells (AD-MSCs) for treating difficult-to-heal skin wounds. Key findings include the validation of a xeno-free enzymatic workflow for keratinocyte extraction, the benefits of using glass as a culture substrate, and the effectiveness of human serum albumin as a transport solution for keratinocytes. The study also reveals that co-culturing keratinocytes and AD-MSCs enhances AD-MSC differentiation towards epidermal cells through miRNA-mediated gene regulations. In vivo experiments using a porcine wound model confirm the regenerative potential of AD-MSCs, highlighting their role in wound healing via miRNA-155 mediated gene regulation. The research provides a comprehensive, EMA-compliant procedure for keratinocyte production and insights into the molecular mechanisms underlying stem cell differentiation and wound healing.