3D Spheroid Culture Synchronizes Heterogeneous MSCs Into an Immunomodulatory Phenotype With Enhanced Anti-Inflammatory Effects

This study demonstrates that culturing mesenchymal stem cells (MSCs) in 3D spheroids enhances their immunomodulatory properties compared to traditional 2D cultures. The 3D spheroid culture synchronizes MSCs into a homogeneous population with increased expression of immunosuppressive factors, such as STC1, TSG6, HGF, and PTGS2, and reduces cell size, minimizing lung entrapment. These 3D MSCs show a greater ability to suppress T lymphocyte proliferation and improve inflammatory lesions in a psoriatic mouse model. Single-cell RNA sequencing revealed that 3D MSCs consist of two major subpopulations with high levels of immunosuppressive factors, unlike the six subpopulations in 2D MSCs. The study suggests that the AP-1 transcription factor complex plays a crucial role in this enhanced immunomodulatory function. These findings indicate that 3D spheroid culture could improve the safety and efficacy of MSC-based therapies for inflammatory diseases.