CILIOGENESIS IN HUMAN BRONCHIAL EPITHELIAL-CELLS CULTURED AT THE AIR-LIQUID INTERFACE
CILIOGENESIS IN HUMAN BRONCHIAL EPITHELIAL-CELLS CULTURED AT THE AIR-LIQUID INTERFACE
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DOI：
10.1165/ajrcmb.10.3.8117445
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发表时间：
1994-03-01
影响因子：
6.4
通讯作者：
PONEC, M
中科院分区：
文献类型：
--
作者：
DEJONG, PM;VANSTERKENBURG, MAJA;PONEC, M
我们研究了改变连续培养的人支气管上皮细胞（HBEC）的培养条件，使支气管上皮细胞与天然上皮相似。从支气管活检中获得细胞，并使用先前描述的方法（体外细胞）进行连续培养。发展生物学1993；29日:379 - 387)。在气液界面处，以去表皮真皮或胶原盘为基质，在含有胎牛血清的培养基中培养第二代及随后的HBEC培养7至31天。扫描电镜和透射电镜显示，无论使用去表皮真皮还是胶原膜，纤毛在7天后发生，成熟时间长达31天。发育中的纤毛透射电镜显示纤维颗粒团块、前纤毛、基体，成熟纤毛的轴突、九对、中心对、径向辐条和纤毛轴的超微结构正常。相比之下，浸泡培养在同一时间内没有纤毛发生的迹象。实验结果表明，通过调节细胞密度、基质的使用和营养补充方式，培养的HBEC暴露在空气中是纤毛发生的必要条件。暴露于空气中的HBEC培养物中纤毛细胞的发育路径与人胎儿气管细胞的分化和成熟模式相似。通过纤维支气管镜获得的人支气管上皮细胞的体外模型为未来研究正常和病理条件下人支气管上皮细胞的功能提供了一个有吸引力的模型。
We present a study on modification of culture conditions in serially cultured human bronchial epithelial cells (HBEC), necessary to achieve bronchial epithelial cells similar to the native epithelium. Cells were obtained from bronchial biopsies and serially cultured using a previously described method (In Vitro Cell. Dev. Biol. 1993; 29A:379-387). At the air-liquid interface, the second and the subsequent passages of HBEC cultures were grown 7 to 31 days, in medium containing fetal calf serum, using de-epidermized dermis or collagen discs as substratum. Scanning and transmission electron microscopy revealed ciliogenesis after 7 days and maturation of the cilia up to 31 days, irrespective of whether de-epidermized dermis or collagen membrane was used. The transmission electron microscopy of the developing cilia showed fibrogranular masses, procentrioles, basal bodies, and in the mature cilia a normal ultrastructure of the axoneme, the nine doublets, the central pair, radial spokes, and dynein arms in the ciliary shaft. In contrast, the submerged cultures showed no signs of ciliogenesis in the same time course. Results of experiments, in which cell seeding density, the substrate used, and the manner of nutrient supplementation were modulated, revealed that the air-exposure of the cultured HBEC is a necessary requirement for the ciliogenesis. The development pathway of ciliated cells in air-exposed HBEC cultures was similar to the differentiation and maturation pattern in human fetal tracheal cells. The in vitro model of human bronchial epithelial cells derived from biopsies obtained by fiberoptic bronchoscopy offers an attractive model for future studies on the function of human bronchial epithelial cells under normal and pathologic conditions.