PDF(Pubmed)
Abstract：
BACKGROUND: AT-MSCs display great immunoregulatory features, making them potential candidates for cell-based therapy. This study aimed to evaluate the \"RBC lysis buffer\" isolation protocol and immunological profiling of the so-obtained AT-MSCs.
METHODS: We established an immune-comparative screening of AT-MSCs throughout in vitro cell expansion (PM, P1, P2, P3, P4) and inflammatory priming regarding the expression of 28 cell-surface markers, 6 cytokines/chemokines, and 10 TLR patterns.
RESULTS: AT-MSCs were highly expandable and sensitive to microenvironment challenges, hereby showing plasticity in distinct expression profiles. Both cell expansion and inflammation differentially modulated the expression profile of CD34, HLA-DR, CD40, CD62L, CD200 and CD155, CD252, CD54, CD58, CD106, CD274 and CD112. Inflammation resulted in a significant increase in the expression of the cytokines IL-6, IL-8, IL-1β, IL-1Ra, CCL5, and TNFα. Depending on the culture conditions, the expression of the TLR pattern was distinctively altered with TLR1-4, TLR7, and TLR10 being increased, whereas TLR6 was downregulated. Protein network and functional enrichment analysis showed that several trophic and immune responses are likely linked to these immunological changes.
CONCLUSIONS: AT-MSCs may sense and actively respond to tissue challenges by modulating distinct and specific pathways to create an appropriate immuno-reparative environment. These mechanisms need to be further characterized to identify and assess a molecular target that can enhance or impede the therapeutic ability of AT-MSCs, which therefore will help improve the quality, safety, and efficacy of the therapeutic strategy.
METHODS: We established an immune-comparative screening of AT-MSCs throughout in vitro cell expansion (PM, P1, P2, P3, P4) and inflammatory priming regarding the expression of 28 cell-surface markers, 6 cytokines/chemokines, and 10 TLR patterns.
RESULTS: AT-MSCs were highly expandable and sensitive to microenvironment challenges, hereby showing plasticity in distinct expression profiles. Both cell expansion and inflammation differentially modulated the expression profile of CD34, HLA-DR, CD40, CD62L, CD200 and CD155, CD252, CD54, CD58, CD106, CD274 and CD112. Inflammation resulted in a significant increase in the expression of the cytokines IL-6, IL-8, IL-1β, IL-1Ra, CCL5, and TNFα. Depending on the culture conditions, the expression of the TLR pattern was distinctively altered with TLR1-4, TLR7, and TLR10 being increased, whereas TLR6 was downregulated. Protein network and functional enrichment analysis showed that several trophic and immune responses are likely linked to these immunological changes.
CONCLUSIONS: AT-MSCs may sense and actively respond to tissue challenges by modulating distinct and specific pathways to create an appropriate immuno-reparative environment. These mechanisms need to be further characterized to identify and assess a molecular target that can enhance or impede the therapeutic ability of AT-MSCs, which therefore will help improve the quality, safety, and efficacy of the therapeutic strategy.
摘要：
背景：AT-MSCs表现出良好的免疫调节功能，使它们成为细胞疗法的潜在候选者。本研究旨在评估“RBC裂解缓冲液”分离方案和如此获得的AT-MSCs的免疫学图谱。
方法：我们在整个体外细胞扩增过程中建立了AT-MSCs的免疫比较筛选（PM，P1，P2，P3，P4）和关于28种细胞表面标志物表达的炎症引发，6细胞因子/趋化因子，和10种TLR模式。
结果：AT-MSCs是高度可扩展的，对微环境挑战敏感，由此在不同的表达谱中显示出可塑性。细胞扩增和炎症差异调节CD34，HLA-DR的表达谱，CD40,CD62L,CD200和CD155、CD252、CD54、CD58、CD106、CD274和CD112。炎症导致细胞因子IL-6，IL-8，IL-1β的表达显着增加，IL-1Ra,CCL5和TNFα。根据培养条件,TLR模式的表达明显改变，TLR1-4，TLR7和TLR10增加，而TLR6下调。蛋白质网络和功能富集分析表明，几种营养和免疫反应可能与这些免疫学变化有关。
结论：AT-MSCs可以通过调节不同的和特定的途径来感知和积极应对组织挑战，以创造适当的免疫修复环境。这些机制需要进一步表征，以鉴定和评估可以增强或阻碍AT-MSC治疗能力的分子靶标。因此，这将有助于提高质量，安全,以及治疗策略的有效性。
方法：我们在整个体外细胞扩增过程中建立了AT-MSCs的免疫比较筛选（PM，P1，P2，P3，P4）和关于28种细胞表面标志物表达的炎症引发，6细胞因子/趋化因子，和10种TLR模式。
结果：AT-MSCs是高度可扩展的，对微环境挑战敏感，由此在不同的表达谱中显示出可塑性。细胞扩增和炎症差异调节CD34，HLA-DR的表达谱，CD40,CD62L,CD200和CD155、CD252、CD54、CD58、CD106、CD274和CD112。炎症导致细胞因子IL-6，IL-8，IL-1β的表达显着增加，IL-1Ra,CCL5和TNFα。根据培养条件,TLR模式的表达明显改变，TLR1-4，TLR7和TLR10增加，而TLR6下调。蛋白质网络和功能富集分析表明，几种营养和免疫反应可能与这些免疫学变化有关。
结论：AT-MSCs可以通过调节不同的和特定的途径来感知和积极应对组织挑战，以创造适当的免疫修复环境。这些机制需要进一步表征，以鉴定和评估可以增强或阻碍AT-MSC治疗能力的分子靶标。因此，这将有助于提高质量，安全,以及治疗策略的有效性。
