Short-term assays for mesenchymal stromal cell immunosuppression of T-lymphocytes

Introduction Trauma patients are susceptible to coagulopathy and dysfunctional immune responses. Mesenchymal stromal cells (MSCs) at the forefront of cellular therapy revolution with profound immunomodulatory, regenerative, therapeutic potential. Routine assays assess immunomodulation activity examine MSC effects on proliferation peripheral blood mononuclear (PBMCs) take 3–7 days. Assays that could be done in a shorter period time would beneficial allow more rapid comparison different donors. The studies presented here focused for suppression mitogen-stimulated PBMC activation frames 24 h or less. Methods Three potential were examined—assays apoptosis focusing caspase activation, phosphatidyl serine externalization (PS+) PBMCs, measurement tumor necrosis factor alpha (TNFα) levels using ELISA methods. All used same initial experimental conditions: cryopreserved PBMCs from 8 10 pooled donors, co-culture without MSCs 96-well plates, stimulation mitogen 2–72 h. Results Suppression activated by incubation was not robust only significant times after Monitoring PS+ live CD3+ CD4+/CD3+ mitogen-activated dose dependent, reproducible, robust, evident earliest point taken, 2 h, although no increase percentage seen time. ability suppress compared favorably two concomitant proliferation, 72 ATP assay, 96 fluorescently labeled protein signal dilution. TNFα release accumulating over However, reliably Discussion Takeaways these as follows: (1) while early measures is 2–6 immunosuppression detected h; (2) PS surrogate assay immunomodulation; (3) detection sensitive