Human bone marrow-derived mesenchymal stem/stromal cells (hMSC) are critical components of tomorrow’s cell-based product and devices. Secretion of biomolecules by hMSC influences many biological processes and is thought to be central to the mechanism of action. Since widespread clinical use of hMSC will be facilitated by frozen storage, cryopreserved hMSC must maintain high levels of biological function upon thaw. To address this critical issue we tested the impact of cryopreservation and thawing on the inducible upregulation of IDO by IFN-g and angiogenic cytokine secretion ((VEGF, HGF, TIMP-1 and -2, FGF2, and IL-8) of hMSC: We hypothesized that cryopreserved hMSC would have diminished immunosuppression response and altered cytokine secretion immediately after thaw compared to hMSC fresh from culture. We compared the biological activity of hMSCs (RoosterBio) from 2 donors either (a) straight out of cryopreservation (THAW), or (b) cells that have been in culture for at least 5 days (FRESH) while controlling for PDL. FRESH or THAW hMSC were plated at 40,000 viable cells/cm2, allowed 4hr attachment, and treated with vehicle or IFN-gamma +/- TNF-alpha for 24 hrs. Induction of IDO activity (immunosuppression) was assayed by measuring kynurenine in the media and cytokine secretion was measured via multiplexed ELISA (Quansys). We report that IDO activity in both FRESH and THAW hMSC was induced by IFN-gamma, and IFN-gamma/TNF-alpha and that the response was not statically different between THAW and FRESH hMSC over multiple experiments. The secreted cytokine profile was also comparable between conditions - opposite of our initial hypothesis based on current literature. These results suggest that cryopreserved hMSC produced using standardized production and cryopreservation processes and solutions can maintain in vitro immunological responsiveness and secreted cytokine activity immediately after thawing. Further work creating negative control is required.