Profiling Mesenchymal Stem Cell Immunomodulation In Vitro

S. Schachtele, D. Galitz, M. Andersen, T. Munn, S. Klingenberg, J. Rivard, C. Hammerbeck, J. Bonnevier, and J. Aho


Mesenchymal stem cells (MSCs) are multipotent adult stem cells found in a variety of tissues, including but not limited to bone marrow, adipose, and umbilical cord. Although MSCs have the ability to differentiate into cell types useful for regenerative medicine, their ability to modulate the immune system has been at the forefront of clinical studies. The mechanisms by which MSCs affect the immune system are not fully understood. This makes it difficult to design specific quality control measures for MSCs used in cell therapy as well as to understand what parameters are the most critical for their immunomodulatory abilities. Defining specific analytical tests and parameters is important for monitoring batch-to-batch variability and to better forecast patient-to-patient efficacy of MSCs used for cell therapy. In this study, we present a workflow to investigate the effects of MSCs on immune cell subset populations. Activated human MSCs were co-cultured with human T cell subtypes (Th1, Th2, Th17, and Treg) that had been differentiated via CellXVivo Differentiation Kits. Co-culture supernates were then analyzed via Proteome Profiler Antibody Arrays to screen for changes in cytokine levels, relative to T cell subsets cultured in the presence of non-activated MSCs. Analytes identified from the screening arrays were then quantitated using Quantikine® ELISA and Luminex® Assays. Additionally, changes in immune cell phenotype were analyzed via flow cytometry. We hypothesize that these tools and techniques allow for a better understanding of the mechanisms behind MSC-mediated immunomodulation and provide methods for routine quality control testing of MSC populations prior to clinical use.