Allogeneic cell-based therapies using adipose tissue-derived stromal cells (ASCs) offer an off-the-shelf alternative to autologous therapy. An underlying assumption is that ASC can modulate the immune response of the recipient. However, in vitro models are required to explore and identify cell interactions and mechanisms of action, to ensure sufficient and sustained effects, and to document these. In this study, we shed light on the effect of ASC manufactured for clinical use on monocyte-derived dendritic cells and an inflammatory microenvironment. ASCs were isolated from healthy voluntary donors, expanded using a human platelet lysate in bioreactors, and cryopreserved as per clinical use. Monocyte-derived dendritic cells were generated by isolation of monocytes and differentiation with GM-CSF and IL-4. Dendritic cells were cocultured with different ratios of ASC and matured with LPS and IFN-γ. Dexamethasone was included as an immunosuppressive control. Dendritic cells were analyzed by flow cytometry for CD11c, CD40, CD80, CD83, CD86, PD-L1, and HLA-DR, and supernatants were analyzed for FGF2, HGF, IL-10, IL-12p70, LIF, MIF, PDGF, PlGF, and IDO. Reduced expression of maturation markers was observed on ASC-treated dendritic cells, while high levels of PD-L1 were maintained. Interestingly, the expression of CD83 was elevated. Escalating ratios of ASC did not affect the concentration of IL-10 considerably, whereas the presence of IL-12 was reduced in a dose-dependent manner. Besides offsetting the IL-12/IL-10 balance, the concentrations of IDO and MIF were elevated in cocultures. Concentrations of FGF2, HGF, LIF, and PIGF were high in ASC cocultures, whereas PDGF was depleted. In a robust coculture model, the addition of ASC to dendritic cells inhibited the dendritic maturation substantially, while inducing a less inflammatory and more tolerogenic milieu. Despite the exposure to dendritic cells and inflammatory stimuli, ASC resulted in supernatants with trophic factors relevant for regeneration. Thus, ASC can perform immunomodulation while providing a regenerative environment.

中文翻译：

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脂肪组织基质细胞诱导高营养环境，同时减少单核细胞衍生树突状细胞的成熟。

使用源自脂肪组织的基质细胞（ASC）的基于异体细胞的疗法提供了自体疗法的现成替代方案。一个基本的假设是ASC可以调节受体的免疫反应。但是，体外需要模型来探索和识别细胞相互作用和作用机制，以确保充分和持续的作用，并记录下来。在这项研究中，我们阐明了临床生产的ASC对单核细胞衍生的树突状细胞和炎性微环境的影响。从健康的自愿供体中分离出ASC，在生物反应器中使用人血小板裂解物扩增ASC，并根据临床用途进行冷冻保存。单核细胞衍生的树突状细胞是通过分离单核细胞并用GM-CSF和IL-4分化而产生的。树突状细胞与不同比例的ASC共培养，并与LPS和IFN- γ一起成熟。包括地塞米松作为免疫抑制对照。通过流式细胞仪分析树突状细胞的CD11c，CD40，CD80，CD83，CD86，PD-L1和HLA-DR，并分析上清液中的FGF2，HGF，IL-10，IL-12p70，LIF，MIF，PDGF， PlGF和IDO。在ASC处理的树突状细胞上观察到成熟标志物的表达降低，而维持了高水平的PD-L1。有趣的是，CD83的表达升高。ASC的升高比例并未显着影响IL-10的浓度，而IL-12的存在以剂量依赖性方式减少。除了抵消IL-12 / IL-10的平衡外，共培养物中IDO和MIF的浓度也升高了。在ASC共培养物中，FGF2，HGF，LIF和PIGF的浓度较高，而PDGF却被耗尽。在强大的共培养模型中，向树突状细胞中添加ASC基本上抑制了树突状成熟，同时诱导了较少的炎症和更多的致耐受性环境。尽管暴露于树突状细胞和炎性刺激，ASC导致上清液中含有与再生相关的营养因子。因此，ASC可以在提供再生环境的同时进行免疫调节。