Culture and Immunomodulation of Equine Muscle-Derived Mesenchymal Stromal Cells: A Comparative Study of Innovative 2D versus 3D Models Using Equine Platelet Lysate.

The research article explores an alternative method for cultivating mesenchymal stromal cells (mdMSCs) from muscle tissue, using equine platelet lysate instead of fetal bovine serum. The study compares this new approach in both 2D and 3D cell culture models.

Introduction and Background

• Mesenchymal stromal cells (mdMSCs), typically derived from muscle tissue, are key players in regenerative medicine because of their ability to modulate immune responses, differentiate into various cell types, and easily harvest.
• However, conventional in vitro expansion practices have several limitations, including ethical concerns and variability between different batches, due to the use of fetal bovine serum (FBS).

Research Objective and Methodology

• This study proposed using 10% equine platelet lysate (ePL), obtained through plasmapheresis, as a substitute for FBS in the cultivation of mdMSCs.
• The researchers explored novel 2D and 3D cell culture models for this purpose, using muscle microbiopsies as the primary source of cells.

Findings and Results

• The study discovered that the heparin concentration had a significant impact on the coagulation of the cell culture medium in 2D cultures, with optimal cell proliferation achieved with a final heparin concentration of 1.44 IU/mL.
• Both the 2D and 3D models affirmed that it’s feasible to expand mdMSCs. The 3D model yielded a higher total number of cells harvested compared to the 2D version.
• Tri-differentiation tests reaffirmed the multipotency of the mdMSCs obtained in the two models, i.e., their ability to differentiate into osteoblasts, chondroblasts, and adipocytes.
• The mdMSCs exhibited classic markers for mesenchymal stem cells (CD-90 and CD-44) and were shown to modulate immune responses effectively. They also inhibited myeloperoxidase (MPO) activity and reduced reactive oxygen species (ROS) activity, with mdMSCs from the 3D model displaying a markedly higher inhibitory effect.

Conclusion and Utility

• The research successfully demonstrates that using 10% equine platelet lysate for the expansion of mdMSCs is not only feasible but also effective using either 2D or 3D cell culture models.
• The strong immunomodulatory activities of harvested mdMSCs highlight their potential in advancing the field of regenerative medicine and cell therapy. This new method also circumvents the issues associated with the use of FBS.