A Simplified 3D-Plasma Culture Method for Generating Minimally Manipulated Autologous Equine Muscle-Derived...

Musculoskeletal pathologies present challenges in athletic horses, often leading to functional impairment. The slow or limited regenerative capacity of bone, joint, and tendon/ligament injuries, coupled with the limitations of conventional treatments, highlights the need for innovative therapies such as ortho-biologics and mesenchymal stem/stroma cells. Traditional 2D cell culture systems with fetal bovine serum (FBS) fail to replicate the complexity of the in vivo environment, whereas 3D cultures more accurately mimic native tissue architecture and cell-cell interactions. This study describes a novel method for isolating muscle-derived progenitor cells in a 3D environment using an autologous plasma-based gel and an innovative cell retrieval solution. The cultured cells exhibit immunomodulatory effects on T lymphocytes, trilineage differentiation potential, and immunophenotypic characteristics consistent with conventional mesenchymal stem/stromal cells. This streamlined 3D culture technique offers a promising platform for generating minimally manipulated autologous cell products tailored for equine regenerative medicine. Key features • Development of a simplified autologous 3D-plasma-based culture method, eliminating the need for fetal bovine serum. • Fully autologous approach offering safer clinical potential for equine regenerative medicine. • Easy and time-efficient method suitable for researchers familiar with basic cell culture techniques.