Institute of Transfusion Medicine, Ulm University Medical Center, Ulm, Germany.
Institute of Clinical Transfusion Medicine and Immunogenetics Ulm, German Red Cross Blood Transfusion Service Baden-Württemberg-Hessen and Ulm University Medical Center, Ulm, Germany.
PLoS One. 2019 May 14;14(5):e0216862. doi: 10.1371/journal.pone.0216862. eCollection 2019.
Polytrauma (PT) is a life-threatening disease and a major global burden of injury. Mesenchymal stromal cells (MSC) might be a therapeutic option for PT patients due to their anti-inflammatory and regenerative potential. We hypothesised that the inflammatory response of MSC is similar after exposure to selected trauma-relevant factors to sera from PT patients (PTS). Therefore, we investigated the effects of a mixture of defined factors, supposed to play a role on MSC in the early phase of PT. Additionally, in a translational approach we investigated the effect of serum from PT patients on MSC in vitro. MSC were incubated with a PT cocktail in physiological (PTCL) and supra-physiological (PTCH) concentrations or PTS. The effect on gene expression and protein secretion of MSC was analysed by RNA sequencing, ELISA and Multiplex assays of cell culture supernatant. Stimulation of MSC with PTCH, PTCL or IL1B led to significant up- or downregulation of 470, 183 and 469 genes compared to unstimulated MSC at 6 h. The intersection of differentially expressed genes in these groups was very high (92% overlap with regard to the PTCL group; treated for 6 h). Cytokine secretion profile of MSC revealed that IL1B mimics the effect of a more complex PT cocktail as well. However, there was only a minor proportion of overlapping differentially expressed genes between the MSC group stimulated with early times of PTS and the MSC group stimulated with PTCH, PTCL and IL1B. In conclusion, the effect of sera from PT patients on MSC activation cannot be simulated by the chosen trauma-relevant factors. Furthermore, we conclude that while IL1B might be useful to prime MSC prior to therapeutic application, it might not be as useful for the in vitro study of functional properties of MSC in the context of PT.
多发伤（PT）是一种危及生命的疾病，也是全球伤害的主要负担。间充质基质细胞（MSC）由于其抗炎和再生潜力，可能成为 PT 患者的治疗选择。我们假设 MSC 在暴露于选定的创伤相关因素后对 PT 患者的血清（PTS）的炎症反应是相似的。因此，我们研究了混合定义因素对 MSC 的影响，这些因素被认为在 PT 的早期阶段对 MSC 起作用。此外，我们采用转化方法研究了 PT 患者的血清对 MSC 的体外作用。MSC 用 PT 鸡尾酒在生理（PTCL）和超生理（PTCH）浓度或 PTS 孵育。通过 RNA 测序、ELISA 和细胞培养上清液的多重分析来分析 MSC 基因表达和蛋白分泌的变化。与未刺激的 MSC 相比，MSC 用 PTCH、PTCL 或 IL1B 刺激 6 小时后，有 470、183 和 469 个基因的表达显著上调或下调。这些组中差异表达基因的交集非常高（与 PTCL 组有 92%的重叠；处理 6 小时）。MSC 细胞因子分泌谱显示，IL1B 模拟了更复杂的 PT 鸡尾酒的作用。然而，在早期 PTS 刺激的 MSC 组和 PTCH、PTCL 和 IL1B 刺激的 MSC 组之间，差异表达基因的重叠比例很小。总之，PT 患者的血清对 MSC 激活的影响不能用选定的创伤相关因素来模拟。此外，我们得出结论，虽然 IL1B 可能有助于在治疗应用前对 MSC 进行预处理，但在 PT 背景下研究 MSC 功能特性的体外研究中，它可能并不那么有用。
