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人血管壁间充质干细胞、真皮成纤维细胞和平滑肌成纤维细胞的分化与可塑性:一项包括成骨潜能超微结构评估的关键比较

Differentiation and plasticity of human vascular wall mesenchymal stem cells, dermal fibroblasts and myofibroblasts: a critical comparison including ultrastructural evaluation of osteogenic potential.

作者信息

Pasanisi Emanuela, Ciavarella Carmen, Valente Sabrina, Ricci Francesca, Pasquinelli Gianandrea

机构信息

Clinical Pathology, Department of Experimental, Diagnostic and Specialty Medicine, DIMES, University of Bologna, University Hospital, S. Orsola, Bologna, Italy.

Immunohaematology and Transfusion Medicine Service, University Hospital, S. Orsola, Bologna, Italy.

出版信息

Ultrastruct Pathol. 2019;43(6):261-272. doi: 10.1080/01913123.2019.1673863. Epub 2019 Oct 7.

DOI:10.1080/01913123.2019.1673863
PMID:31589079
Abstract

Mesenchymal stem cells (MSCs) share many properties with other tissue stromal cells, including cell morphology, immunophenotype, differentiation and immunologic properties. In this study, we compared the immunophenotype and the differentiation potential of human vascular wall mesenchymal stem cells (hVW-MSCs) with those of human dermal fibroblasts and myofibroblasts. Cell morphology and surface markers were evaluated by immunofluorescence and flow cytometry; functional assays for immunomodulation, angiogenesis, adipogenesis and osteogenesis were performed, together with the mRNA analysis of the critical differentiation genes. hVW-MSCs, dermal fibroblasts and myofibroblasts were all negative to CD34, whereas the expression of CD44 stemness marker was more intense in hVW-MSCs. As expected, hVW-MSC plasticity was wide and the angiogenic, adipogenic, osteogenic features were confirmed. Fibroblasts were the less effective in terms of immunomodulation, angiogenesis and adipogenic differentiation; differently from fibroblasts, the myofibroblasts showed a poor angiogenic commitment. The mineralization assay was positive in all the three cell types, but ultrastructure interestingly evidenced differential osteogenic patterns among them. Our study supports the higher anti-inflammatory and wound healing repair features of hVW-MSCs, in comparison to the other stromal cells investigated. Moreover, we underline the importance of ultrastructure for investigating the specific osteogenic pattern for each cell type.

摘要

间充质干细胞(MSCs)与其他组织基质细胞具有许多共同特性,包括细胞形态、免疫表型、分化和免疫特性。在本研究中,我们比较了人血管壁间充质干细胞(hVW-MSCs)与人真皮成纤维细胞和肌成纤维细胞的免疫表型及分化潜能。通过免疫荧光和流式细胞术评估细胞形态和表面标志物;进行免疫调节、血管生成、脂肪生成和成骨的功能测定,以及关键分化基因的mRNA分析。hVW-MSCs、真皮成纤维细胞和肌成纤维细胞对CD34均呈阴性,而CD44干性标志物在hVW-MSCs中的表达更强。正如预期的那样,hVW-MSC的可塑性广泛,血管生成、脂肪生成和成骨特征得到证实。成纤维细胞在免疫调节、血管生成和脂肪生成分化方面效果较差;与成纤维细胞不同,肌成纤维细胞的血管生成能力较弱。矿化试验在所有三种细胞类型中均呈阳性,但超微结构有趣地显示出它们之间不同的成骨模式。我们的研究支持hVW-MSCs与其他所研究的基质细胞相比具有更高的抗炎和伤口愈合修复特性。此外,我们强调了超微结构对于研究每种细胞类型特定成骨模式的重要性。

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