Figure S1. Flow cytometric gates for the evaluation and collection of B lineage cells from the bone marrow of 8-week C57BL/6 mice. Table S1. CDR-H3 sequences obtained from wild-type C57BL/6 bone marrow B lineage cells Table S2. CDR-H3 sequences obtained from C57BL/6 IgHa.ΔD-iD congenic bone marrow mature, recirculating
B cells. “
“There is emerging interest in the application of mesenchymal stem cells (MSC) for the prevention and treatment of autoimmune diseases, graft-versus-host disease and allograft rejection. It is, however, unknown how inflammatory conditions affect phenotype and function of MSC. Adipose tissue-derived mesenchymal Proteasome inhibitor stem cells (ASC) were cultured with alloactivated peripheral blood mononuclear cells (PBMC) (mixed lymphocyte reaction: MLR), with proinflammatory cytokines [interferon (IFN)-γ, tumour necrosis factor (TNF)-α and interleukin (IL)-6] or under control conditions,
and their full genome expression and function examined. Proinflammatory cytokines mainly increased indoleamine-2,3-dioxygenase expression, whereas ASC cultured with MLR showed increased expression of COX-2, involved in prostaglandin E2 production. Both conditions had a stimulatory, but differential, BMN 673 mouse effect on the expression of proinflammatory cytokines and chemokines, while the expression of fibrotic factors was decreased only in response to proinflammatory cytokines. Functional analysis demonstrated that inflammatory conditions affected morphology and proliferation of ASC, while their differentiation capacity and production of trophic factors was unaffected. The immunosuppressive capacity
of ASC was enhanced strongly under inflammatory conditions. In conclusion, ASC showed enhanced immunosuppressive capacity under inflammatory conditions, while their differentiation capacity was preserved. Therefore, Tobramycin in vitro preconditioning provides ASC with improved properties for immediate clinical immune therapy. Mesenchymal stem cells (MSC) are found in a variety of tissues, including bone marrow, skin and adipose tissue [1–3] and can be expanded easily in vitro. MSC are thought to have tissue regenerative properties, in the first place via their multi-lineage differentiation capacity  and, perhaps more importantly, via the secretion of trophic factors that may activate local progenitor cells . In addition, MSC have potent immunomodulatory capacity. They inhibit the proliferation of T cells [5,6] and inhibit dendritic cell maturation [7,8]. These properties make MSC promising for a diversity of clinical applications; for example, for the prevention and treatment of autoimmune diseases and bone marrow rejection. Recently, interest has developed in the use of MSC in solid organ transplantation [9,10]. These conditions are associated with an inflammatory milieu.