Metabolomic Profiling of Equine Cord Blood Mesenchymal Stromal Cells in Both Early and Late Stages of Culture

Abstract Introduction: Equine Mesenchymal Stromal Cells (MSC) hold great potential as a future form of cellular therapy. Fetal Bovine Serum (FBS) is used as a supplement MSC culture, but challenges with its use have initiated a desire to create a defined media to expand these cells. MSCs have shown great variability in species, tissue source and passage number. Due to this, to effectively create a replacement for FBS we must better understand how it meets the metabolic needs for each type of MSC. Objectives: The goal of this study is to determine key pathways and differences in pathways between the early and late passages of equine cord blood MSCs. Methods: This study utilized metabolomics to give a snapshot of the metabolism in the long-term culture of equine cord blood MSCs (eCB-MSC) by comparing the profile of plain culture media to spend media. Results: We found two significantly different metabolites between the early and late passage of eCB-MSC, alpha-ketoglutaric acid (p = 0.019) and creatine (p = 0.012). As well, the metabolic results allowed us to perform an enrichment analysis to assess which pathway(s) were most relevant. Conclusion: The two metabolites suggest a difference in metabolism between the early and late passage reflecting different cellular priorities. Insights into the unique metabolism of these cells and how the requirement of the cell differs between the early and late passage may allow the formulation a serum free media tailored to the metabolic needs of eCB-MSC.