Tancharoen Waleephan, Aungsuchawan Sirinda, Pothacharoen Peraphan, Markmee Runchana, Narakornsak Suteera, Kieodee Junjira, Boonma Nonglak, Tasuya Witoon
Department of Anatomy, Faculty of Medicine, Chiang Mai University, Chiang Mai, Thailand.
Department of Anatomy, Faculty of Medicine, Chiang Mai University, Chiang Mai, Thailand.
Acta Histochem. 2017 Mar;119(2):113-121. doi: 10.1016/j.acthis.2016.11.009. Epub 2016 Dec 22.
Endothelial dysfunction is a principle feature of vascular-related disease. Endothelial cells have been acquired for the purposes of the restoration of damaged tissue in therapeutic angiogenesis. However, their use is limited by expansion capacity and the small amount of cells that are obtained. Human amniotic fluid mesenchymal stem cells (hAF-MSCs) are considered an important source for vascular tissue engineering. In this study, hAF-MSCs were characterized and then induced in order to differentiate into the endothelial-like cells. Human amniotic fluid cells (hAFCs) were obtained from amniocentesis at the second trimester of gestation. The cells were characterized as mesenchymal stem cells by flow cytometry. The results showed that the cells were positive for mesenchymal stem cell markers CD44, CD73, CD90 and HLA-ABC, and negative for CD31, Amniotic fluid stem cells marker: CD117, anti-human fibroblasts, HLA-DR and hematopoietic differentiation markers CD34 and CD45. The hAF-MSCs were differentiated into endothelial cells under the induction of vascular endothelial growth factor (VEGF) and analyzed for the expression of the endothelial-specific markers and function. The expression of the endothelial-specific markers was determined by reverse transcriptase-quantitative PCR (RT-qPCR), while immunofluorescent analysis demonstrated that the induced hAF-MSCs expressed von Willebrand factor (vWF), vascular endothelial growth factor receptor 2 (VEGFR2), CD31 and endothelial nitric oxide synthase (eNOS). The network formation assay showed that the induced hAF-MSCs formed partial networks. All results indicated that hAF-MSCs have the potential to be differentiated into endothelial-like cells, while human amniotic fluid might be a suitable source of MSCs for vascularized tissue engineering.
内皮功能障碍是血管相关疾病的主要特征。在治疗性血管生成中，已获取内皮细胞用于修复受损组织。然而，它们的应用受到扩增能力和所获细胞数量少的限制。人羊水间充质干细胞（hAF-MSCs）被认为是血管组织工程的重要来源。在本研究中，对hAF-MSCs进行了表征，然后诱导其分化为内皮样细胞。人羊水细胞（hAFCs）取自妊娠中期的羊膜穿刺术。通过流式细胞术将这些细胞表征为间充质干细胞。结果显示，这些细胞对间充质干细胞标志物CD44、CD73、CD90和HLA-ABC呈阳性，而对CD31、羊水干细胞标志物：CD117、抗人成纤维细胞、HLA-DR以及造血分化标志物CD34和CD45呈阴性。hAF-MSCs在血管内皮生长因子（VEGF）的诱导下分化为内皮细胞，并对内皮特异性标志物的表达和功能进行分析。通过逆转录定量PCR（RT-qPCR）测定内皮特异性标志物的表达，而免疫荧光分析表明，诱导后的hAF-MSCs表达血管性血友病因子（vWF）、血管内皮生长因子受体2（VEGFR2）、CD31和内皮型一氧化氮合酶（eNOS）。网络形成试验表明，诱导后的hAF-MSCs形成了部分网络。所有结果表明，hAF-MSCs具有分化为内皮样细胞的潜力，而人羊水可能是用于血管化组织工程的间充质干细胞的合适来源。
