Kress Taylor C, Barris Candee T, Kennard Simone, Belin de Chantemèle Eric J
Vascular Biology Center, Medical College of Georgia at Augusta University, Augusta, GA, USA.
Department of Medicine (Cardiology), Medical College of Georgia at Augusta University, Augusta, GA, USA.
Bio Protoc. 2025 Sep 5;15(17):e5440. doi: 10.21769/BioProtoc.5440.
Cardiovascular disease, the current leading cause of death worldwide, is a multifactorial disorder that involves a strong contribution of both the innate and adaptive immune systems. Overactivation of the immune system and inappropriate secretion of pro-inflammatory cytokines lead to vascular impairments and the development of cardiovascular disorders, including hypertension, atherosclerosis, and peripheral artery disease. Lymphocytes, macrophages, and dendritic cells can all secrete pro-inflammatory cytokines. This makes it challenging to isolate a specific subset of immune cells, particularly cytokines, and their contribution to vascular dysfunction remains difficult to elucidate. To solve this problem, our laboratory has developed the novel "immune cell-aorta" co-culture system described herein. This experimental protocol enables investigators to isolate an immune cell of interest and identify the cytokine(s) at the origin of vascular alterations. Key features • Novel ex vivo approach combining the culture of one population of immune cells with blood vessels. • No direct contact between the cells and the blood vessels; the model enables studying the role of immune cell-derived factors or cytokines on vascular function. • Blood vessels can subsequently be used for functional (wire/pressure myography), molecular (western blot, quantitative real-time RT-PCR), and histological studies.
心血管疾病是目前全球主要的死亡原因，它是一种多因素疾病，涉及先天性和适应性免疫系统的强烈作用。免疫系统的过度激活和促炎细胞因子的不适当分泌会导致血管损伤以及心血管疾病的发生，包括高血压、动脉粥样硬化和外周动脉疾病。淋巴细胞、巨噬细胞和树突状细胞都能分泌促炎细胞因子。这使得分离特定的免疫细胞亚群，尤其是细胞因子变得具有挑战性，并且它们对血管功能障碍的作用仍难以阐明。为了解决这个问题，我们实验室开发了本文所述的新型“免疫细胞-主动脉”共培养系统。该实验方案使研究人员能够分离出感兴趣的免疫细胞，并确定血管改变起源处的细胞因子。关键特性•将一群免疫细胞与血管培养相结合的新型体外方法。•细胞与血管之间无直接接触；该模型能够研究免疫细胞衍生因子或细胞因子对血管功能的作用。•血管随后可用于功能（线/压力肌动描记法）、分子（蛋白质印迹法、定量实时逆转录聚合酶链反应）和组织学研究。
