Development of an in vitro human nasal epithelial (HNE) cell model
Development of an in vitro human nasal epithelial (HNE) cell model
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DOI：
10.1016/0378-4274(96)03720-4
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发表时间：
1996-11-01
影响因子：
3.5
通讯作者：
Aufderheide, M
中科院分区：
文献类型：
--
作者：
Schmidt, D;Hubsch, U;Aufderheide, M
上呼吸道主要参与对空气中物质的反应。因此，人鼻组织的原代培养可以为评估环境污染物引起的毒性或炎症过程提供应用模型。通过蛋白酶消化从鼻组织活检中分离出 HNE 细胞，并在无血清的补充激素的培养基中培养。当在空气-液体界面的多孔膜上生长时，与保持浸没的细胞相比，在两周内发育出更高比例的纤毛细胞。胶原凝胶基质上的浸没生长的培养物保持立方上皮样形态并且细胞角蛋白染色呈阳性。鼻细胞的原代培养物表现出显着的酸性磷酸酶活性。第二代HNE细胞在用颗粒物质处理后表现出显着的酶反应活性。与用天然柴油机尾气颗粒处理的细胞相比，用甲苯提取的柴油机尾气颗粒处理的细胞中溶酶体标记酶酸性磷酸酶的诱导更强。天然柴油颗粒的非特异性酯酶活性增加了 7 倍以上，而提取颗粒诱导的酯酶活性与对照组相比相对较低。
The upper respiratory tract is mainly involved in reactions to airborne substances. So the primary culture of human nasal tissue may provide an application model for assessment of toxic or inflammatory processes caused by environmental pollutants. HNE cells were isolated from nasal tissue biopsies by protease digestion and cultured in serum-free hormone-supplemented medium. When grown on porous membranes at the air-liquid interface, a higher portion of ciliated cells developed within 2 weeks compared with cells maintained submerged. Submerged grown cultures on collagen gel matrix maintained a cuboidal epithelial-like morphology and stained positive for cytokeratin. The primary cultures of nasal cells exhibited a marked activity for acid phosphatase. Second passage HNE cells exhibited marked enzymatic reactivity upon treatment with particulate matter. Induction of the lysosomal marker enzyme acid phosphatase was stronger in cells treated with toluene-extracted diesel exhaust particles compared with cells treated with native diesel exhaust particles. Non-specific esterase activity was increased more than 7-fold by native diesel particles, whereas the induction of esterase activity by extracted particles was relatively low compared with the control group.