Ibrahim Asmaa Q, Abdullah Mohammed S, Ahram Mamoun, Abdalla Shtaywy
Department of Biological Sciences, School of Science, The University of Jordan, Amman 11942, Jordan.
Department of Physiology and Biochemistry, School of Medicine, The University of Jordan, Amman 11942, Jordan.
Methods Protoc. 2023 May 26;6(3):54. doi: 10.3390/mps6030054.
Vascular smooth muscle cells (VSMCs) and vascular endothelial cells are key participants in the pathogenesis of atherosclerosis. Human umbilical vein endothelial cells (HUVECs) and VSMCs are useful models to design therapeutic strategies for many cardiovascular diseases (CVDs). However, procuring a VSMC cell line by researchers, to model atherosclerosis, for example, is impeded by time and cost limitations, as well as by many other logistic problems in many countries.
This article describes a protocol for the quick and cheap isolation of VSMCs from human umbilical cords using a mechanical and enzymatic method. This VSMC protocol yields a confluent primary culture that could be obtained within 10 days and sub-cultured for 8-10 passages. The isolated cells are characterized by their morphology and the expression of mRNA of marker proteins analyzed by reverse transcription polymerase chain reaction (RT-qPCR).
The protocol described herein for the isolation of VSMCs from human umbilical cords is easy and is time- and cost-efficient. Isolated cells are useful models for understanding the mechanisms underlying many pathophysiological conditions.
血管平滑肌细胞（VSMC）和血管内皮细胞是动脉粥样硬化发病机制的关键参与者。人脐静脉内皮细胞（HUVEC）和VSMC是设计许多心血管疾病（CVD）治疗策略的有用模型。然而，例如，研究人员获取VSMC细胞系以模拟动脉粥样硬化时，会受到时间和成本限制以及许多国家的其他诸多后勤问题的阻碍。
本文描述了一种使用机械和酶法从人脐带中快速、廉价分离VSMC的方法。该VSMC方法可产生融合的原代培养物，可在10天内获得，并可传代培养8 - 10代。通过形态学以及逆转录聚合酶链反应（RT - qPCR）分析的标记蛋白mRNA表达对分离的细胞进行表征。
本文所述的从人脐带中分离VSMC的方法简便，且具有时间和成本效益。分离的细胞是理解许多病理生理状况潜在机制的有用模型。
