Human Co- and Triple-Culture Model of the Alveolar-Capillary Barrier on a Basement Membrane Mimic
Human Co- and Triple-Culture Model of the Alveolar-Capillary Barrier on a Basement Membrane Mimic
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DOI：
10.1089/ten.tec.2018.0087
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发表时间：
2018-09-01
影响因子：
3
通讯作者：
Kirkpatrick, C. James
中科院分区：
文献类型：
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作者：
Dohle, Eva;Singh, Smriti;Kirkpatrick, C. James
开发类似肺泡-毛细血管屏障的体外模型可能是研究肺部生理学以及肺部对感染或暴露于纳米颗粒后的免疫反应的非常有益的工具。本研究是基于在基膜模拟物上开发的体外肺泡屏障，该基膜模拟物由超薄纳米纤维网状物组成，该网状物由生物可吸收聚（epsilon-己内酯）静电纺丝制成。作为细胞组分，采用类似肺泡上皮细胞的NCI H441和内皮细胞系ISO-HAS-1进行双极共培养实验，总培养期为14天。除了免疫组织化学和免疫荧光研究外，还研究了体外模型系统的经上皮电阻（TER）和运输能力。肺泡屏障功能可以在基底膜模拟物的双极共培养系统中清楚地确定，因为TER在双极培养过程中增加。此外，为了首次深入了解体外可能的肺部炎症反应，该共培养模型通过人白血病单核细胞系（THP-1）进一步扩展。这种三重培养系统能够充分保持双极共培养的屏障特性，从而使这种由基底膜上的上皮细胞、内皮细胞和免疫细胞组成的体外模型为组织工程的进一步研究奠定了良好的基础。
The development of an in vitro model resembling the alveolar-capillary barrier might be a highly beneficial tool to study lung physiology as well as the immune response of the lung to infection or after exposure to nanoparticles. This study is based on an in vitro alveolar barrier developed on a basement membrane mimic, composed of ultrathin nanofiber meshes generated via electrospinning using bioresorbable poly(epsilon-caprolactone). As cellular components, NCI H441, resembling the alveolar epithelial cells, and ISO-HAS-1, an endothelial cell line, were used to perform bipolar coculture experiments for a total cultivation period of 14 days. In addition to immunohistochemical and immunofluorescent studies, transepithelial electrical resistance (TER) and transport capabilities of the in vitro model system were investigated. Alveolar barrier function could be clearly determined for the postulated bipolar coculture system on the basement membrane mimic, since TER increased during the course of bipolar cultivation. Furthermore, to gain first insights into possible lung inflammatory reactions in vitro, this coculture model was further expanded by a human leukemia monocyte cell line (THP-1). This triple-culture system was able to maintain adequately the barrier properties of the bipolar coculture, thus making this in vitro model consisting of epithelial, endothelial, and immune cells on a basement membrane mimic a promising basis for further studies in tissue engineering.