Oosterhoff Loes A, Kruitwagen Hedwig S, van Wolferen Monique E, van Balkom Bas W M, Mokry Michal, Lansu Nico, van den Dungen Noortje A M, Penning Louis C, Spanjersberg Talitha C F, de Graaf Johannes W, Veenendaal Tomas, Zomerdijk Flin, Fledderus Joost O, Spee Bart, van Steenbeek Frank G
Department of Clinical Sciences of Companion Animals, Faculty of Veterinary Medicine, Utrecht University, Utrecht, Netherlands.
Nephrology and Hypertension, Division of Internal Medicine and Dermatology, University Medical Center Utrecht, Utrecht, Netherlands.
Front Physiol. 2019 Feb 12;10:101. doi: 10.3389/fphys.2019.00101. eCollection 2019.
Vasculature performs a critical function in tissue homeostasis, supply of oxygen and nutrients, and the removal of metabolic waste products. Vascular problems are implicated in a large variety of pathologies and accurate models resembling native vasculature are of great importance. Unfortunately, existing models do not sufficiently reflect their counterpart. The complexity of vasculature requires the examination of multiple cell types including endothelial cells (ECs) and vascular smooth muscle cells (VSMCs), as well as vessel location in the body from which they originate. The use of canine blood vessels provides a way to study vasculature with similar vessel size and physiology compared to human vasculature. We report an isolation procedure that provides the possibility to isolate both the endothelial and smooth muscle cells from the same vessels simultaneously, enabling new opportunities in investigating vasculature behavior. Canine primary ECs and VSMCs were isolated from the vena cava, vena porta and aorta. All tissue sources were derived from three donors for accurate comparison and to reduce inter-animal variation. The isolation and purification of the two distinct cell types was confirmed by morphology, gene- and protein-expression and function. As both cell types can be derived from the same vessel, this approach allows accurate modeling of vascular diseases and can also be used more widely, for example, in vascular bioreactors and tissue engineering designs. Additionally, we identified several new genes that were highly expressed in canine ECs, which may become candidate genes for novel EC markers. In addition, we observed transcriptional and functional differences between arterial- and venous-derived endothelium. Further exploration of the transcriptome and physiology of arteriovenous differentiation of primary cells may have important implications for a better understanding of the fundamental behavior of the vasculature and pathogenesis of vascular disease.
血管系统在组织稳态、氧气和营养物质供应以及代谢废物清除方面发挥着关键作用。血管问题与多种病理状况相关，因此，能够准确模拟天然血管系统的模型至关重要。不幸的是，现有的模型并不能充分反映其对应物。血管系统的复杂性要求对多种细胞类型进行研究，包括内皮细胞（ECs）和血管平滑肌细胞（VSMCs），以及它们在体内的起源位置。使用犬类血管提供了一种研究血管系统的方法，其血管大小和生理机能与人类血管系统相似。我们报告了一种分离程序，该程序提供了从同一血管中同时分离内皮细胞和平滑肌细胞的可能性，为研究血管行为带来了新的机遇。犬类原代内皮细胞和平滑肌细胞从腔静脉、门静脉和主动脉中分离出来。所有组织来源均来自三个供体，以进行准确比较并减少动物个体间的差异。通过形态学、基因和蛋白质表达以及功能，证实了两种不同细胞类型的分离和纯化。由于这两种细胞类型都可以从同一血管中获得，这种方法能够准确模拟血管疾病，还可更广泛地应用，例如用于血管生物反应器和组织工程设计。此外，我们鉴定出了几个在犬类内皮细胞中高表达的新基因，这些基因可能成为新型内皮细胞标志物的候选基因。此外，我们观察到动脉源性和静脉源性内皮细胞之间的转录和功能差异。进一步探索原代细胞动静脉分化的转录组和生理学，可能对更好地理解血管系统的基本行为和血管疾病的发病机制具有重要意义。
