Ciliary and secretory differentiation of normal human middle ear epithelial cells
Ciliary and secretory differentiation of normal human middle ear epithelial cells
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DOI：
10.1080/000164802753648141
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发表时间：
2002-04-01
影响因子：
1.4
通讯作者：
Yoon, JH
中科院分区：
文献类型：
--
作者：
Choi, JY;Kim, CH;Yoon, JH
然而，最近的技术进步现在允许正常人类中耳上皮细胞（NHMEE）的连续培养。这些细胞的纤毛分化尚未实现。本研究的目的是建立一个培养系统，使连续培养的NHMEE细胞分化为纤毛细胞。如果纤毛细胞发育，在整个培养过程中测量纤毛细胞和分泌细胞的百分比。我们还以时间依赖性的方式检测了粘蛋白和溶菌酶分泌水平及其mrna。我们收集了外观正常的人中耳粘膜，并在酶解后进行了连续培养。这些细胞在空气-液界面（ALI）培养系统中培养2年。7. 14. 合流后21天和28天。纤毛发生通常在融合后16天开始，免疫组织化学染色检测纤毛细胞百分比！随着时间的推移Lip增加到a，最大可达10.6%，但在融合后的第14天培养期间，分泌细胞的百分比保持稳定在约40%的吞吐量。粘蛋白和溶酶酶分泌量。经点印迹法测定，其含量显著增加后保持稳定。粘蛋白基因5B （MUC5B）的表达水平。MUC8和溶菌酶随培养时间的延长而增加。其中MUC8在融合后第28天显著增加；MUC5AC mRNA水平在融合后第14天达到峰值。综上所述，ALI培养系统可以诱导NHMEE细胞的纤毛分化。我们的研究还表明，分泌功能的发育早于纤毛发生，MUC5B和MUC8 mrna的表达随着纤毛分化而增加。
Recent technical advances now permit the serial culture of normal human middle ear epithelial (NHMEE) cells, However. the ciliary differentiation of these cells has not been achieved. The purpose of this study was to C establish a Culture S system in order to differentiate serially cultured NHMEE cells into ciliated cells. If ciliated cells developed, the percentages of ciliated cells and secretory cells were measured throughout the duration of culture. We also examined the levels of mucin and lysozyme secretion and their mRNAs in a time-dependent manner, Human middle ear mucosa with a normal appearance was harvested and serially cultured after enzymatic disaggregation. These cells were Cultured in In air liquid interface (ALI) culture system for 2. 7. 14. 21 and 28 days after confluence. Ciliogenesis usually began 16 IS days after confluence, The percentage of ciliated cells detected by means of immunohistochemical staining! increased Over time Lip to a, maximum of 10.6% but the percentage of secretors cells remained stable at approximate to 40% throughput the duration Of Culture By Day 14 after confluence. the amounts of mucin and lysosyme secretion. its measured by dot-blotting analysis, had increased significantly and then remained stable. The expression levels of mucin gene 5B (MUC5B). MUC8 and lysozyme increased with the duration of culture. MUC8 in particular showed a dramatic increase on Day 28 after confluence:c. In contrast, the level of MUC5AC mRNA peaked On Day 14 after confluence. and then decreased, in conclusions, ciliary differentiation of NHMEE cells can be induced using an ALI culture system, Our study also suggest, that secretory function develops earlier than ciliogenesis, and that the expressions of MUC5B and MUC8 mRNAs increase as a function of differentiation.