Phenotypic, morphological, and metabolic characterization of vascular-spheres from human vascular mesenchymal stem cells,Microscopy Research and Technique

当前位置： X-MOL 学术 › Microsc. Res. Tech. › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

Phenotypic, morphological, and metabolic characterization of vascular-spheres from human vascular mesenchymal stem cells
Microscopy Research and Technique ( IF 2.0 ) Pub Date : 2021-08-27 , DOI: 10.1002/jemt.23918
Sabrina Valente ₁ , Carmen Ciavarella ₁ , Anna Hernández-Aguilera _{2,

3} , Fernández-Arroyo Salvador _{2,

3} , Marina Buzzi ₄ , Jorge Joven _{2,

3} , Gianandrea Pasquinelli _{1,

5}

Affiliation

The ability to form spheroids under non-adherent conditions is a well-known property of human mesenchymal stem cells (hMSCs), in addition to stemness and multilineage differentiation features. In the present study, we tested the ability of hMSCs isolated from the vascular wall (hVW-MSCs) to grow as spheres, and provide a characterization of this 3D model. hVW-MSCs were isolated from femoral arteries through enzymatic digestion. Spheres were obtained using ultra-low attachment and hanging drop methods. Immunophenotype and pluripotent genes (SOX-2, OCT-4, NANOG) were analyzed by immunocytochemistry and real-time PCR, respectively. Spheres histological and ultrastructural architecture were examined. Cell viability and proliferative capacity were measured using LIVE/DEATH assay and ki-67 proliferation marker. Metabolomic profile was obtained with liquid chromatography–mass spectrometry. In 2D, hVW-MSCs were spindle-shaped, expressed mesenchymal antigens, and displayed mesengenic potential. 3D cultures of hVW-MSCs were CD44⁺, CD105^low, CD90^low, exhibited a low propensity to enter the cell cycle as indicated by low percentage of ki-67 expression and accumulated intermediate metabolites pointing to slowed metabolism. The 3D model of hVW-MSCs exhibits stemness, dormancy and slow metabolism, typically observed in stem cell niches. This culture strategy can represent an accurate model to investigate hMSCs features for future clinical applications in the vascular field.

中文翻译：

人类血管间充质干细胞血管球的表型、形态学和代谢特征

除了干性和多向分化特征外，在非粘附条件下形成球体的能力是人间充质干细胞 (hMSC) 的一个众所周知的特性。在本研究中，我们测试了从血管壁（hVW-MSCs）分离的 hMSCs 生长为球体的能力，并提供了该 3D 模型的表征。通过酶消化从股动脉中分离出 hVW-MSC。使用超低附着和悬滴法获得球体。分别通过免疫细胞化学和实时 PCR 分析免疫表型和多能基因（SOX-2、OCT-4、NANOG）。检查球体组织学和超微结构结构。使用 LIVE/DEATH 测定和 ki-67 增殖标记物测量细胞活力和增殖能力。通过液相色谱-质谱法获得代谢组学特征。在 2D 中，hVW-MSCs 呈纺锤形，表达间充质抗原，并显示出间生潜能。hVW-MSCs 的 3D 培养物是 CD44⁺ , CD105^低, CD90^低, 表现出进入细胞周期的低倾向，如 ki-67 表达的低百分比和表明代谢减慢的中间代谢物积累所示。hVW-MSCs 的 3D 模型表现出干性、休眠和缓慢的新陈代谢，通常在干细胞壁龛中观察到。这种培养策略可以代表一个准确的模型来研究 hMSCs 的特征，以供将来在血管领域的临床应用中使用。

更新日期：2021-08-27

点击分享查看原文

点击收藏

公开下载

阅读更多本刊最新论文本刊介绍/投稿指南11