Zhao Huaning, Darden Jordan, Chappell John C
Center for Heart and Regenerative Medicine, Virginia Tech Carilion Research Institute, Roanoke, VA, USA.
Department of Biomedical Engineering and Mechanics, Virginia Polytechnic Institute and State University, Blacksburg, VA, USA.
Microcirculation. 2018 Jul;25(5):e12461. doi: 10.1111/micc.12461. Epub 2018 Jun 7.
Pericytes are specialized perivascular cells embedded within the basement membrane. These cells envelope the abluminal surface of endothelial cells and promote microvessel homeostasis. Recent discoveries of unique pericyte functions, particularly in neural tissues, underscore the need for overcoming existing challenges in establishing a functionally validated pericyte cell line. Here, we present methodologies for addressing these challenges as well as an embryonic pericyte cell line for use with in vitro and ex vivo experimental models.
We isolated an enriched population of NG2:DsRed+ pericytes from E12.5 mice. This pericyte cell line was compared to MEFs with respect to gene expression, cell morphology and migration, and engagement with endothelial cells during junction stabilization and angiogenesis.
NG2+ pericytes displayed gene expression patterns, cell morphology, and 2D migration behaviors distinct from MEFs. In three different vessel formation models, pericytes from this line migrated to and incorporated into developing vessels. When co-cultured with HUVECs, these pericytes stimulated more robust VE-Cadherin junctions between HUVECs as compared to MEFs, as well as contributed to HUVEC organization into primitive vascular structures.
Our data support use of this pericyte cell line in a broad range of models to further understand pericyte functionality during normal and pathological conditions.
周细胞是嵌入基底膜内的特殊血管周围细胞。这些细胞包绕内皮细胞的无腔面并促进微血管稳态。近期对周细胞独特功能的发现,尤其是在神经组织中的功能,凸显了克服在建立功能验证的周细胞系方面现有挑战的必要性。在此,我们介绍应对这些挑战的方法以及一种用于体外和离体实验模型的胚胎周细胞系。
我们从E12.5小鼠中分离出富集的NG2:DsRed+周细胞群体。将该周细胞系在基因表达、细胞形态和迁移以及在连接稳定和血管生成过程中与内皮细胞的相互作用方面与小鼠胚胎成纤维细胞(MEFs)进行比较。
NG2+周细胞表现出与MEFs不同的基因表达模式、细胞形态和二维迁移行为。在三种不同的血管形成模型中,该细胞系的周细胞迁移至并整合到发育中的血管中。当与人类脐静脉内皮细胞(HUVECs)共培养时,与MEFs相比,这些周细胞刺激HUVECs之间形成更强健的VE-钙黏蛋白连接,并且有助于将HUVECs组织成原始血管结构。
我们的数据支持在广泛的模型中使用该周细胞系,以进一步了解正常和病理条件下周细胞的功能。