Primary Human Lung Pericytes Support and Stabilize In Vitro Perfusable Microvessels
Primary Human Lung Pericytes Support and Stabilize In Vitro Perfusable Microvessels
复制标题
DOI：
10.1089/ten.tea.2014.0545
复制
发表时间：
2015-08-01
影响因子：
4.1
通讯作者：
Geiser, Thomas
中科院分区：
文献类型：
--
作者：
Bichsel, Colette A.;Hall, Sean R. R.;Geiser, Thomas
血管的形成是复杂的组织特异性过程，其在发育过程、伤口愈合、癌症进展、纤维化和其他病理学中起关键作用。为了研究肺中的血管发生和血管重塑，我们开发了一种体外微血管模型，该模型在三维结构、可及性、功能性和细胞类型方面密切模仿人肺微血管。使用流式细胞术分离和表征来自远端气道的人周细胞。为了评估它们在正常微血管生成中的作用，将肺周细胞混合在纤维蛋白凝胶中，并与原代内皮细胞（人脐带静脉内皮细胞）一起接种到明确定义的微区室中。3-5天内形成面积为3.1 mm（2）的通畅微血管，并稳定长达14天。来自肺周细胞的可溶性信号是建立灌注性所必需的，周细胞向内皮微血管迁移。透射电镜和免疫细胞化学芯片证实细胞间以粘附和紧密连接的形式进行通讯，以及分泌基底膜。周细胞与内皮细胞直接共培养使微血管通透性降低一个数量级，从17.8 × 10（-6）cm/s降低到2.0 × 10（-6）cm/s，并导致血管直径明显变小且变化较小。苯肾上腺素给药后，在衬有周细胞的微血管中观察到血管收缩，但仅在内皮微血管中未观察到。还用人肺微血管内皮细胞和肺周细胞产生可灌注的微血管。因此，人肺周细胞显示出在体外微血管系统中对微血管形态、渗透性、血管收缩和长期稳定性具有显著影响。这种仿生平台为在生理相关的微血管环境中测试患者来源的细胞的功能和相互作用开辟了新的可能性。
The formation of blood vessels is a complex tissue-specific process that plays a pivotal role during developmental processes, in wound healing, cancer progression, fibrosis, and other pathologies. To study vasculogenesis and vascular remodeling in the context of the lung, we developed an in vitro microvascular model that closely mimics the human lung microvasculature in terms of three-dimensional architecture, accessibility, functionality, and cell types. Human pericytes from the distal airway were isolated and characterized using flow cytometry. To assess their role in the generation of normal microvessels, lung pericytes were mixed in fibrin gel and seeded into well-defined microcompartments together with primary endothelial cells (human umbilical cord vein endothelial cells). Patent microvessels covering an area of 3.1mm(2) formed within 3-5 days and were stable for up to 14 days. Soluble signals from the lung pericytes were necessary to establish perfusability, and pericytes migrated toward endothelial microvessels. Cell-cell communication in the form of adherens and tight junctions, as well as secretion of basement membrane were confirmed using transmission electron microscopy and immunocytochemistry on chip. Direct coculture of pericytes with endothelial cells decreased the microvascular permeability by one order of magnitude from 17.8x10(-6) to 2.0x10(-6) cm/s and led to vessels with significantly smaller and less variable diameter. Upon phenylephrine administration, vasoconstriction was observed in microvessels lined with pericytes, but not in endothelial microvessels only. Perfusable microvessels were also generated with human lung microvascular endothelial cells and lung pericytes. Human lung pericytes were thus shown to have a prominent influence on microvascular morphology, permeability, vasoconstriction, and long-term stability in an in vitro microvascular system. This biomimetic platform opens new possibilities to test functions and interactions of patient-derived cells in a physiologically relevant microvascular setting.