Enhancing autologous Equine Therapy: Innovative 3D Culture Method for Muscle-derived Progenitor Cells (mdP-Cells)

Abstract Musculoskeletal pathologies pose significant challenges in athletic horses, often leading to loss of function. The limited healing capacity of bone, joint, and tendon/ligament injuries, combined with the inefficacy of conventional treatments, underscores the need for innovative therapeutic strategies such as Mesenchymal Stem/Stroma Cells (MSCs). Conventional cell culture methods typically employ two-dimensional (2D) culture systems with adherent cells using fetal bovine serum. However, three-dimensional (3D) cell culture offers a more physiologically relevant milieu, closely resembling native tissue architecture and in vivo cell-cell interactions. This study introduces a novel approach for isolating muscle-derived progenitor cells in a 3D environment, utilizing an autologous plasma-based gel and an innovative cell retrieval solution. The muscle-derived progenitor cells cultured using this method exhibited immunomodulatory properties on lymphocyte T and trilineage differentiation and immunophenotyping akin to traditional MSCs. This simplified 3D culture technique holds promise for generating minimally manipulated autologous cell products tailored for musculoskeletal applications.