In vitro self-assembly of human pericyte-supported endothelial microvessels in three-dimensional coculture: a simple model for interrogating endothelial-pericyte interactions.
In vitro self-assembly of human pericyte-supported endothelial microvessels in three-dimensional coculture: a simple model for interrogating endothelial-pericyte interactions.
复制标题
三维共培养中人周细胞支持的内皮微血管的体外自组装：用于询问内皮-周细胞相互作用的简单模型。
DOI：
10.1159/000353303
复制
发表时间：
2013
影响因子：
1.7
通讯作者：
Pober,JS
中科院分区：
文献类型：
--
作者：
Waters,JP;Kluger,MS;Graham,M;Chang,WG;Bradley,JR;Pober,JS
我们描述了一种在三维(3-D)蛋白质基质中共培养宏微血管人内皮细胞(ECs)和周细胞(PC)的方法，导致由PC投资的管状ECs。为了防止三维培养中内皮细胞的凋亡性死亡，转导人脐静脉或真皮微血管内皮细胞表达抗凋亡蛋白Bcl2。为了防止PC介导的凝胶收缩，胶原-纤维连接蛋白凝胶在聚乙醇酸非织造基质中聚合。在最初的24-48小时内，仅含EC的凝胶自发形成管束，通过空泡化形成管腔；这样的血管网络维持长达7天。在EC-PC共文化中，PC被招募到EC网络中。血管内皮细胞支架植入血管内皮细胞后，管腔直径减小，血管网分支程度增加。在该体系中，EC和PC之间形成了聚乙二醇式和插座式连接，但没有观察到染料转移，这表明缝隙连接的形成。这个简单的系统可以用来分析3-D几何结构中EC和PC之间的双向信号。
We describe a method for coculture of macro-or microvascular human endothelial cells (ECs) and pericytes (PCs) within a 3-dimensional (3-D) protein matrix resulting in lumenized EC cords invested by PCs. To prevent apoptotic cell death of ECs in 3-D culture, human umbilical vein or dermal microvascular ECs were transduced to express the antiapoptotic protein Bcl-2. To prevent PC-mediated gel contraction, the collagen-fibronectin gel was polymerized within a polyglycolic acid nonwoven matrix. Over the first 24-48 h, EC-only gels spontaneously formed cords that developed lumens via vacuolization; such vascular networks were maintained for up to 7 days. In EC-PC cocultures, PCs were recruited to the EC networks. PC investment of EC cords both limited the lumen diameter and increased the degree of vascular network arborization. Peg and socket junctions formed between ECs and PCs in this system, but dye transfer, indicative of gap junction formation, was not observed. This simple system can be used to analyze bidirectional signals between ECs and PCs in a 3-D geometry.