Abstract
Regulatory dendritic cells (DCregs) represent a potential therapeutic tool for assessing a variety of immune overreaction conditions; however, current approaches for generating DCregs for therapeutic purposes are limited. We attempted to generate and characterize DCregs from murine induced pluripotent stem (iPS) cells. The iPS cells co-cultured with OP9 cells displayed mesodermally differentiated flat colonies. GM-CSF drove most of the colonies exhibiting a differentiated morphology. Thereafter, cells became morphologically heterologous under the effects of TGF-β and IL-10. Most of the floating cells developed an irregular shape with areas of protrusion. The generated iPS-DCregs demonstrated high CD11b/c and low CD40, CD80, CD86 and MHC-II expressions with a high antigen uptake ability and poor T-cell stimulatory function. Importantly, iPS-DCregs showed immune responsiveness regulation effects both in vitro and in vivo and the ability to generate regulatory T-cells in vitro. Our result illustrates a feasible approach for generating functional DCregs from murine iPS cells.
Highlights
Regulatory dendritic cells (DCregs) represent a potential therapeutic tool for assessing a variety of immune overreaction conditions; current approaches for generating DCregs for therapeutic purposes are limited
We describe a novel approach for generating a high number of functional DCregs from induced pluripotent stem cells for downregulating the immune response. iPS cells were first reported by Takahashi and Yamanaka in 2006 and can be produced from various types of somatic cells via reprogramming by Yamanaka factors (Oct[4], Sox[2], Klf[4] and c-Myc)22. iPS cells are very similar to embryonic stem (ES) cells in many respects, including gene expression patterns and pluripotent characteristics; they are not restricted by the same ethical concerns as ES cells
The present study investigated iPS-MEF-Ng-38C-2, a previously established murine iPS cell clone, for its capacity to differentiate into functional DCregs
Summary
Regulatory dendritic cells (DCregs) represent a potential therapeutic tool for assessing a variety of immune overreaction conditions; current approaches for generating DCregs for therapeutic purposes are limited. Hong et al found that constructed adenoviral vector coding suppressor of cytokine signaling (SOCS)-1 can efficiently increase the SOCS-1 gene expression in BM-DCcons and induce DCreg generation, possessing the therapeutic potential to prevent rejection in patients undergoing organ transplantation[11] Summarizing these reports, the in vitro generation of DCregs has been achieved using many manipulations, such as the following three methods: 1) physiological mediators, including anti-inflammatory cytokines, such as IL-10, TGF-b1 and vascular endothelial growth factor (VEGF)12–14; 2) pharmacological agents, including anti-inflammatory agents (such as aspirin), cyclic adenosine monophosphate www.nature.com/scientificreports (cAMP) inducers (prostaglandin E2, histamine, b2 agonists, neuropeptides [calcitonin-gene-related peptide and vasoactive intestinal peptide]), vitamin D3 and immunosuppressive drugs (corticosteroids, cyclosporin A, rapamycin, deoxyspergualin and mycophenolate mofetil (MMF))[15,16,17,18]; and 3) genetic engineering of molecules, such as co-stimulatory molecules and cytokines, that can be transferred through viral or nonviral delivery systems or manipulated by selective gene silencing [antisense oligodeoxynucleotides (ODNs) and small interfering RNAs (siRNAs)]19–21. We investigated the use of OP9 stromal cells as feeder cells accompanying the combination of exogenous GM-CSF and the antiinflammatory cytokines IL-10 and TGF-b as culture conditions to generate DCregs from murine iPS cells (iPS-DCreg) and characterized the cells using morphological, related gene-expression and functional analyses
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