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Characterization of Bone Marrow-Derived Dendritic Cells Developed in Serum-Free Media and their Ability to Prevent Type 1 Diabetes in Nonobese Diabetic Mice

Ben M Looney, Anna V Chernatynskaya, Michael J Clare-Salzler and Chang-Qing Xia

Dendritic cells (DC) have been investigated as a cell-based therapy for Type 1 Diabetes (T1D). BM-DC expanded ex vivo with GM-CSF and IL-4 is typically cultured with fetal bovine serum (FBS). The effect of FBS on NOD BM-DC has not been extensively studied. In the present study we compare BM-DC generated in serumfree culture media (X-VIVO20; FBS-) with BM-DC generated in media containing 10% FBS (RPMI1640/10%FBS; FBS+). We show that FBS- BM-DC display a phenotype and cytokine-producing profile distinct from FBS+ BMDC. Additionally, compared to FBS+ BM-DC, we show evidence of an altered Th cell response induced by FBSBM- DC. Finally, we demonstrate that only FBS- BM-DC prevent the onset of T1D and induce increased levels of CD4+Foxp3+ regulatory T cells as well as a long-lasting β cell-specific T cell response. This study indicates that serum-free media generates a more tolerogenic BM-DC capable of preventing T1D in the NOD mice.