Forskolin Enhances Urokinase Plasminogen Activator Secretion and Angiogenic Activity of Xeno‐Free Cultures of Human Adipose Tissue‐Derived Stem Cells

Abstract Adipose tissue‐derived stem cells (ASCs) hold significant potential for treating various clinical conditions. To enhance their regenerative properties, ASCs can be chemically stimulated using various in vitro protocols. However, unsatisfactory results persist, partly due to the relatively costly long‐term methods. Furthermore, current culturing techniques often rely on the use of xenogenic fetal bovine serum that can be immunogenic, limiting clinical translations. To facilitate clinical translation of ASCs‐derived therapeutics, the effect of different stimulation protocols on human ASCs cultured in a xeno‐free medium (PRIME‐XV MSC Expansion XSFM) is investigated. The xeno‐free medium was supplemented with stimulants (forskolin (FSK), basic fibroblast growth factor, platelet‐derived growth factor‐AA, neuregulin‐1) in combinations or individually. Stimulation for 72 h in FSK alone, or together with the growth factors, enhanced the production of urokinase plasminogen activator (uPA), a serine protease involved in tissue remodeling processes. Conditioned medium derived from stimulated ASCs enhanced in vitro angiogenesis and endothelial cells migration. This study shows that pro‐angiogenic responses in human ASCs can be enhanced with a defined short stimulation protocol using a xeno‐free medium. The protocol, using readily available manufacturing cell therapy grade molecules, may boost the regenerative properties of ASCs secretome which could enhance their efficacy in clinical treatments.