Paudel Puja, McDonald Fiona, Fronius Martin
Department of Physiology, University of Otago, Dunedin, New Zealand.
HeartOtago, University of Otago, Dunedin, New Zealand.
Methods Mol Biol. 2025;2894:21-34. doi: 10.1007/978-1-0716-4342-6_3.
Endothelial cells are a major contributor to cardiovascular diseases. Studying their function and how they influence pathological processes remains an ongoing area of research. Although primary endothelial cells might be readily available from animals, translating results from these studies can be challenging due to species-dependent differences. A common source of human endothelial cells is human umbilical vein endothelial cells, but these cells might not represent the variety of endothelial cells from various vascular beds. Here we describe a protocol for the isolation and cultivation of endothelial cells from human internal mammary artery remains. These remains are commonly available from coronary artery bypass graft surgeries. The described tissue explant method combined with a magnetic sorting process provided a relatively high yield of endothelial cells that were subsequently passaged and used for studies involving mRNA and protein expression analyses, as well as fluorescence-based immunohistochemistry and patch-clamp electrophysiology. This protocol may help to extend the repertoire of studying human endothelial cells to understand their role and function in health and disease.
内皮细胞是心血管疾病的主要促成因素。研究它们的功能以及它们如何影响病理过程仍是一个正在进行的研究领域。尽管原代内皮细胞可能很容易从动物身上获得，但由于物种依赖性差异，将这些研究结果进行转化可能具有挑战性。人内皮细胞的一个常见来源是人类脐静脉内皮细胞，但这些细胞可能无法代表来自各种血管床的内皮细胞的多样性。在此，我们描述了一种从人乳内动脉残余物中分离和培养内皮细胞的方案。这些残余物通常可从冠状动脉搭桥手术中获得。所描述的组织外植体方法与磁分选过程相结合，提供了相对高产的内皮细胞，这些细胞随后传代并用于涉及mRNA和蛋白质表达分析以及基于荧光的免疫组织化学和膜片钳电生理学的研究。该方案可能有助于扩展研究人类内皮细胞的方法，以了解它们在健康和疾病中的作用和功能。
