Purpose. This study is aimed at investigating the phenotype, differentiation potential, immunomodulatory properties, and responsiveness of saphenous vein vessel wall-derived mesenchymal stromal cells (SV-MSCs) to various TLR ligands and proinflammatory cytokines, as well as comparing their features to those of their bone marrow-derived counterparts (BM-MSCs). Methods. SV-MSCs were isolated by enzymatic digestion of the saphenous vein vessel wall. Phenotype analysis was carried out by flow cytometry and microscopy, whereas adipogenic, chondrogenic, and osteogenic differentiation potentials were tested in in vitro assays. For comparative analysis, the expression of different stemness, proliferation, and differentiation-related genes was determined by Affymetrix gene array. To compare the immunomodulatory properties of SV-MSCs and BM-MSCs, mixed lymphocyte reaction was applied. To investigate their responses to various activating stimuli, MSCs were treated with TLR ligands (LPS, PolyI:C) or proinflammatory cytokines (TNFα, IL-1β, IFNγ), and the expression of various early innate immune response-related genes was assessed by qPCR, while secretion of selected cytokines and chemokines was measured by ELISA. Results. The isolated SV-MSCs were able to differentiate into bone, fat, and cartilage cells/direction in vitro. SV-MSCs expressed the most important MSC markers (CD29, CD44, CD73, CD90, and CD105) and shared almost identical phenotypic characteristics with BM-MSCs. Their gene expression pattern and activation pathways were close to those of BM-MSCs. SV-MSCs showed better immunosuppressive activity inhibiting phytohemagglutinin-induced T lymphocyte proliferation in vitro than BM-MSCs. Cellular responses to treatments mimicking inflammatory conditions were comparable in the bone marrow- and saphenous vein-derived MSCs. Namely, similar to BM-MSCs, SV-MSCs secreted increased amount of IL-6 and IL-8 after 12- or 24-hour treatment with LPS, PolyI:C, TNFα, or IL-1β, compared to untreated controls. Interestingly, a different CXCL-10/IP-10 secretion pattern could be observed under inflammatory conditions in the two types of MSCs. Conclusion. Based on our results, cells isolated from saphenous vein vessel wall fulfilled the ISCT’s (International Society for Cellular Therapy) criteria for multipotent mesenchymal stromal cells, and no significant differences in the phenotype, gene expression pattern, and responsiveness to inflammatory stimuli could be observed between BM-MSCs and SV-MSCs, while the latter cells have more potent immunosuppressive activity in vitro. Further functional assays have to be performed to reveal whether SV-MSCs could be useful for certain regenerative therapeutic applications or tissue engineering purposes.

中文翻译：

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血管壁来源的间充质基质细胞与骨髓来源的基质细胞具有相似的分化潜能和免疫调节特性

目的。本研究旨在研究隐静脉血管壁来源的间充质基质细胞 (SV-MSCs) 对各种 TLR 配体和促炎细胞因子的表型、分化潜能、免疫调节特性和反应性，并将它们的特征与它们的特征进行比较。骨髓来源的对应物（BM-MSCs）。方法。通过大隐静脉血管壁的酶消化分离SV-MSC。通过流式细胞术和显微镜进行表型分析，而在体外测试脂肪形成、软骨形成和成骨分化潜能化验。为了比较分析，通过Affymetrix基因阵列测定不同干性、增殖和分化相关基因的表达。为了比较 SV-MSCs 和 BM-MSCs 的免疫调节特性，应用了混合淋巴细胞反应。为了研究它们对各种激活刺激的反应，MSCs 用 TLR 配体（LPS、PolyI:C）或促炎细胞因子（TNFα 、IL - 1β、IFNγ ）处理，以及各种早期先天免疫反应相关基因的表达通过 qPCR 评估，而通过 ELISA 测量选定的细胞因子和趋化因子的分泌。结果。分离的 SV-MSCs 能够在体外分化成骨、脂肪和软骨细胞/方向. SV-MSCs 表达最重要的 MSC 标志物（CD29、CD44、CD73、CD90 和 CD105），并与 BM-MSCs 具有几乎相同的表型特征。它们的基因表达模式和激活途径与BM-MSCs接近。SV-MSCs在体外抑制植物血凝素诱导的 T 淋巴细胞增殖的免疫抑制活性优于 BM-MSCs。在骨髓和大隐静脉衍生的 MSCs 中，细胞对模拟炎症状况的治疗反应相当。即，与 BM-MSCs 相似，SV-MSCs 在用 LPS、PolyI:C、TNF α或 IL-1 β处理 12 或 24 小时后分泌增加量的 IL-6 和 IL-8，与未处理的对照相比。有趣的是，在两种类型的 MSCs 的炎症条件下，可以观察到不同的 CXCL-10/IP-10 分泌模式。结论。根据我们的结果，从大隐静脉血管壁分离的细胞符合 ISCT（国际细胞治疗学会）对多能间充质基质细胞的标准，并且在表型、基因表达模式和对炎症刺激的反应性方面没有观察到显着差异。 BM-MSCs 和 SV-MSCs，而后者细胞在体外具有更强的免疫抑制活性。必须进行进一步的功能测定以揭示 SV-MSC 是否可用于某些再生治疗应用或组织工程目的。