Rothen-Rutishauser Barbara M, Kiama Stephen G, Gehr Peter
Institute of Anatomy, Division of Histology, University of Bern, Baltzerstrasse 2, CH-3000 Bern 9, Switzerland.
Am J Respir Cell Mol Biol. 2005 Apr;32(4):281-9. doi: 10.1165/rcmb.2004-0187OC. Epub 2005 Jan 7.
A novel triple co-culture model of the human airway barrier was designed to simulate the cellular part of the air-blood barrier of the respiratory tract represented by macrophages, epithelial cells, and dendritic cells. When epithelial cells (A549 cells) were grown on filter inserts with pores of 3.0 mum in diameter in a two-chamber system, they formed monolayers with polarization into apical and basolateral domains. The epithelial cell cultures were then supplemented with human blood monocyte-derived macrophages and dendritic cells on the apical and basal aspect, respectively. The single-cell cultures as well as the triple co-cultures were characterized in terms of a number of typical features, for example, morphology of cell types, integrity of epithelial layer, and expression of specific cell surface markers (CD14 for macrophages and CD86 for dendritic cells). The interplay of epithelial cells with macrophages and dendritic cells during the uptake of polystyrene particles (1 mum in diameter) was investigated with confocal laser scanning and conventional transmission electron microscopy. Particles were found in all three cell types, although dendritic cells were not directly exposed to the particles. More investigations are needed to understand the translocation pathway.
设计了一种新型的人气道屏障三重共培养模型，以模拟由巨噬细胞、上皮细胞和树突状细胞代表的呼吸道气血屏障的细胞部分。当上皮细胞（A549细胞）在两室系统中直径为3.0μm的滤膜小室上生长时，它们形成具有顶端和基底外侧结构域极化的单层。然后分别在顶端和基底方面用人血单核细胞衍生的巨噬细胞和树突状细胞补充上皮细胞培养物。单细胞培养物和三重共培养物根据许多典型特征进行表征，例如细胞类型的形态、上皮层的完整性以及特定细胞表面标志物（巨噬细胞的CD14和树突状细胞的CD86）的表达。用共聚焦激光扫描和传统透射电子显微镜研究了上皮细胞与巨噬细胞和树突状细胞在摄取聚苯乙烯颗粒（直径1μm）过程中的相互作用。尽管树突状细胞未直接暴露于颗粒，但在所有三种细胞类型中均发现了颗粒。需要更多的研究来了解转运途径。
