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Abstract
Mesenchymal stromal/stem cells (MSCs) are multipotent adult stem cells which show immunomod-ulation capacity and have been widely used in clinical trials. Despite their immunomodulationpotential, to date, there is no FDA-approved MSC therapy available. MSCs show heterogeneityat different levels, including donor-donor variation, tissue-tissue variation and cell subpopulationvariation. Therefore, it is difficult to predict MSC immunomodulation effects as well as predictoutcome after MSC therapy. The lack of predictive markers which can indicate MSC immunomod-ulation effects limits its use in clinic.In this dissertation, I developed new methods to characterize MSC immunomodulation effects usingmetabolomics. I first investigated the cellular metabolomics and media cytokines to predict MSCimmunomodulation effects. The composite functional scores were generated from T cell suppres-sion assay and IDO activity results. These composite scores were then predicted using partial leastsquares (PLSR) model using metabolomics and cytokines profiles. Several metabolites includingphosphocreatine, NN-dimethylglycine, asparagine, myo-inositol and serine were found importantin predicting composite functional score, therefore, can be used as surrogates to predict MSC im-munomodulation effects. These methods, although can accurate predict MSC function, however,are all destructive to cells. Non-destructive, in-process characterization of MSCs is needed. Timecourse monitoring of culture media can closely reflect cell phenotype and can be used to predictMSC function. I found that metabolites change rate of first three days culturing in culture mediacan be used to predict MSC immunomodulation effects. These metabolites include leucine, thre-onine and alanine. Based on these findings, pathway analysis was also used to reveal some of theunderlying mechanisms. The pathway analysis showed that cells consume aspartate to generateasparagine, meanwhile, secrete alanine and glycine to culture meida. Based on these findings, I alsoaim to validate the predictive markers using additional nine bone marrow-derived MSC lines. Thebottleneck of MSC research is the application in cell manufacturing. I also discussed the applicationof media monitoring using bioreactors and bench-top NMR in industrial setting.