Institute of Biomedical and Clinical Science, Peninsula Medical School, University of Exeter, St Luke's Campus, Exeter, Devon, UK.
Microcirculation. 2011 Nov;18(8):635-45. doi: 10.1111/j.1549-8719.2011.00128.x.
Despite an increasing research demand for human microvascular endothelial cells, isolation of primary endothelial cells from human tissue remains difficult. The omentum, a highly vascular visceral adipose tissue, could provide an excellent source of these cells.
A reliable method to isolate HOMECs has been developed. It consists of initial enzymatic digestion (to deplete cell contaminants), followed by further digestion, selective filtration, and immunoselection using Dynabeads coated with CD31 antibody. Cultures were characterized for expression of endothelial cell markers and their ability to undergo VEGF-dependent in vitro tube structure formation.
Omental-derived cultures of microvascular endothelial cells were achieved with <5% contamination of other cell types. The endothelial origin of cells was confirmed by the constitutive expression of a range of vascular endothelial markers (CD31, CD105, vWF) and internalization of DiI-AcLDL. Furthermore, cultures were negative for lymphatic endothelial markers, underwent in vitro angiogenesis, and exhibited typical endothelial morphology.
This isolation method produces homogeneous HOMEC cultures that can be maintained in vitro for at least six passages without loss of cellular features characterizing endothelial cells.
尽管对人类微血管内皮细胞的研究需求不断增加，但从人体组织中分离原发性内皮细胞仍然很困难。大网膜是一种高度血管化的内脏脂肪组织，可以作为这些细胞的绝佳来源。
开发了一种可靠的分离 HOMEC 的方法。它包括初始酶消化（以耗尽细胞污染物），然后进一步消化、选择性过滤和使用包被有 CD31 抗体的 Dynabeads 进行免疫选择。培养物的特征在于表达内皮细胞标志物及其在 VEGF 依赖性体外管结构形成中的能力。
大网膜来源的微血管内皮细胞培养物中其他细胞类型的污染<5%。细胞的内皮起源通过一系列血管内皮标志物（CD31、CD105、vWF）的组成型表达和 DiI-AcLDL 的内化得到证实。此外，培养物对淋巴管内皮标志物呈阴性，进行体外血管生成，并表现出典型的内皮形态。
这种分离方法可产生同质的 HOMEC 培养物，可在体外至少传代 6 代而不丧失内皮细胞的特征。
Zotero 插件