Ciliogenesis in submersion and suspension cultures of human nasal epithelial cells
Ciliogenesis in submersion and suspension cultures of human nasal epithelial cells
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DOI：
10.1007/s00405-002-0562-y
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发表时间：
2003-07-01
影响因子：
2.6
通讯作者：
Walger, M
中科院分区：
文献类型：
--
作者：
Neugebauer, P;Endepols, H;Walger, M
人鼻呼吸道细胞在深层培养中失去纤毛。本研究比较了细胞外基质（ECM）分子的基底层对纤毛发生在浸没细胞培养无ECM悬浮培养。鼻黏膜上皮细胞的培养以呼吸道黏膜为主要来源。对于浸没培养物，将酶促分离的细胞接种在致死辐射（Co-60，60戈伊）的鼠3 T3饲养成纤维细胞层上或ECM包被的培养瓶上。对于悬浮培养物，细胞接种后将烧瓶旋转3天。在ECM包被的烧瓶中，上皮细胞附着和汇合得到促进，并且总是比饲养层上的培养物好得多。呼吸道细胞在浸没培养的前5周失去纤毛。当用ECM分子涂覆塑料培养皿时，在5-6周后观察到新的、活跃跳动的纤毛的发生。与3 T3-成纤维细胞一起在未涂覆的塑料培养皿上生长的细胞显示无纤毛发生。悬浮培养的上皮细胞球体在培养第1周纤毛脱落，1-2周后纤毛再生。我们的研究结果表明，ECM分子不是唯一的信号纤毛分化的呼吸细胞在体外，因为悬浮培养是ECM免费。然而，在浸没的细胞培养物中ECM分子的存在促进了具有高密度立方上皮细胞的接种上皮细胞的附着和早期汇合。这些立方细胞的特殊形状和细胞间的紧密接触可能是诱导终末分化和纤毛发生的重要信号之一。
Human nasal respiratory cells lose cilia in submerged cultures. This study compares the effect of extracellular matrix (ECM) molecules of the basal lamina on ciliogenesis in submerged cell cultures to ECM-free suspension cultures. Respiratory mucosa of nasal turbinates was the routine source for the cultures of nasal epithelial cells. For the submersion cultures, enzymatically isolated cells were seeded either on a layer of lethally irradiated (Co-60, 60 Gy) murine 3T3-feeder fibroblasts or on an ECM-coated culture flask. For suspension cultures, the flasks were rotated for 3 days after cell seeding. In ECM-coated flasks, epithelial cell attachment and confluence was promoted and always much better than in cultures on a feeder layer. Respiratory cells lost cilia during the first 5 weeks in submerged cultures. Genesis of new, actively beating cilia was seen after 5-6 weeks when plastic culture dishes were coated with ECM molecules. Cells grown on uncoated plastic dishes together with 3T3-fibroblasts showed no ciliogenesis. Spheroids of epithelial cells in suspension cultures lost cilia during the 1st week and developed new cilia after 1-2 weeks in vitro. Our results suggest that ECM molecules are not the only signal for ciliary differentiation of respiratory cells in vitro, because suspension cultures are ECM free. However, the presence of ECM molecules in submerged cell cultures promotes the attachment and early confluence of seeded epithelial cells with a high density of cuboidal epithelial cells. The specific cellular shape and intense intercellular contact of these cuboidal cells may be among the most important signals inducing terminal differentiation and ciliogenesis.