Airway branching morphogenesis in three dimensional culture.
Airway branching morphogenesis in three dimensional culture.
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DOI：
10.1186/1465-9921-11-162
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发表时间：
2010-11-25
影响因子：
5.8
通讯作者：
Magnusson MK
中科院分区：
文献类型：
--
作者：
Franzdóttir SR;Axelsson IT;Arason AJ;Baldursson O;Gudjonsson T;Magnusson MK
肺从胎儿消化道发育而来，在此过程中上皮细胞侵入血管丰富的基质，称为分支形态发生。在器官发生中，内皮细胞已被证明对于器官结构的形态发生和维持是重要的。本研究的目的是通过建立一个三维（3D）的共培养模型，肺上皮细胞培养在富含内皮细胞的基质中，在体外重现人肺的形态发生。我们使用了我们实验室最近开发的人支气管上皮细胞系（VA 10）。该细胞系细胞系保持主要的基底细胞表型，表达p63和其他基底标志物，如细胞角蛋白-5和-14。在这里，我们将VA 10与人脐静脉内皮细胞（HUVEC）一起培养，以模拟肺发育过程中这些细胞类型之间的密切相互作用。形态发生和分化通过相差显微镜、免疫染色和共聚焦成像进行监测。我们发现，在与内皮细胞共培养时，VA 10细胞产生细支气管肺泡样结构，表明肺上皮分支是由内皮细胞的存在促进的。VA 10衍生的上皮结构显示出各种复杂的分支模式，并显示出部分肺泡II型分化与前表面活性剂-C表达。通过免疫染色证实分支VA 10集落的上皮来源。这些细支气管肺泡样结构相对于细胞-基质界面处的整合素表达被极化。可溶性因子介导内皮诱导的分支。此外，成纤维细胞生长因子受体-2（FGFR-2）和sprougty-2在分支结构的生长尖端表达，并且分支被FGFR-小分子抑制剂SU 5402抑制。在这项研究中，我们表明，人肺上皮细胞系可以诱导内皮细胞形成分支支气管肺泡样结构的三维培养。这种新的人类气道形态发生模型可用于研究人类肺发育中的关键事件，并表明内皮在促进气道上皮分支中的支持作用。
Lungs develop from the fetal digestive tract where epithelium invades the vascular rich stroma in a process called branching morphogenesis. In organogenesis, endothelial cells have been shown to be important for morphogenesis and the maintenance of organ structure. The aim of this study was to recapitulate human lung morphogenesis in vitro by establishing a three dimensional (3D) co-culture model where lung epithelial cells were cultured in endothelial-rich stroma. We used a human bronchial epithelial cell line (VA10) recently developed in our laboratory. This cell line cell line maintains a predominant basal cell phenotype, expressing p63 and other basal markers such as cytokeratin-5 and -14. Here, we cultured VA10 with human umbilical vein endothelial cells (HUVECs), to mimic the close interaction between these cell types during lung development. Morphogenesis and differentiation was monitored by phase contrast microscopy, immunostainings and confocal imaging. We found that in co-culture with endothelial cells, the VA10 cells generated bronchioalveolar like structures, suggesting that lung epithelial branching is facilitated by the presence of endothelial cells. The VA10 derived epithelial structures display various complex patterns of branching and show partial alveolar type-II differentiation with pro-Surfactant-C expression. The epithelial origin of the branching VA10 colonies was confirmed by immunostaining. These bronchioalveolar-like structures were polarized with respect to integrin expression at the cell-matrix interface. The endothelial-induced branching was mediated by soluble factors. Furthermore, fibroblast growth factor receptor-2 (FGFR-2) and sprouty-2 were expressed at the growing tips of the branching structures and the branching was inhibited by the FGFR-small molecule inhibitor SU5402. In this study we show that a human lung epithelial cell line can be induced by endothelial cells to form branching bronchioalveolar-like structures in 3-D culture. This novel model of human airway morphogenesis can be used to study critical events in human lung development and suggests a supportive role for the endothelium in promoting branching of airway epithelium.
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影响因子：
56.9
作者：
Lammert, E;Cleaver, O;Melton, D
通讯作者：
Melton, D
影响因子：
56.9
作者：
Matsumoto, K;Yoshitomi, H;Zaret, KS
通讯作者：
Zaret, KS
影响因子：
13.6
作者：
Rønnov-Jessen L;Bissell MJ
通讯作者：
Bissell MJ
影响因子：
4.9
作者：
Asgrimsson, V;Gudjonsson, T;Baldursson, O
通讯作者：
Baldursson, O
DOI：
10.1164/ajrccm.157.6.9707011
发表时间：
1998-06-01
影响因子：
24.7
作者：
Boers, JE;Ambergen, AW;Thunnissen, FBJM
通讯作者：
Thunnissen, FBJM