人鼻腔纤毛上皮细胞的浸没培养

发布时间：2018-06-21 13:36

  本文选题：人 + 鼻腔　； 参考：《临床耳鼻咽喉头颈外科杂志》2014年12期

【摘要】：目的:浸没培养法培养人鼻腔纤毛上皮细胞,为纤毛相关研究及经鼻药物安全性评价等研究提供可靠的细胞学模型。方法:采用低温酶消化法,浸没培养人鼻腔纤毛上皮细胞,倒置相差显微镜观察细胞生长状况,扫描电镜及免疫细胞化学方法观察细胞融合及纤毛分化状态,高速数字化显微视频成像系统检测纤毛摆动频率。结果:①相差显微镜下,纤毛细胞数量逐渐增加,至7~10d达到高峰后逐渐减少,纤毛细胞存活时间维持在14~21d;②细胞培养第7天,扫描电镜可见鼻腔上皮细胞表面覆盖纤毛或微绒毛,杯状细胞及无纤毛柱状上皮细胞相间排列;③细胞培养第7天,Ⅳ型β-微管蛋白、闭合小环蛋白-1免疫荧光结果显示细胞融合及纤毛分化良好,纤毛细胞比例可达20%~30%;④培养第7、14、21天纤毛摆动基础频率分别为(10.73±2.15)Hz、(9.92±1.97)Hz、(10.30±2.11)Hz,无明显统计学差异;⑤外源性刺激剂100μmol/L ATP可明显增加纤毛摆动频率。结论:应用酶消化法浸没培养人鼻腔纤毛上皮细胞,细胞融合及纤毛分化状态良好,纤毛摆动活跃,对外源性刺激反应灵敏,是应用于纤毛相关研究及经鼻药物安全性评价等研究的一个较为理想的细胞模型。
[Abstract]:Objective: to culture human nasal ciliated epithelial cells by immersion culture, and to provide a reliable cytological model for the study of cilium and safety evaluation of transnasal drugs. Methods: human nasal ciliated epithelial cells were incubated by low temperature enzyme digestion. The growth status of cells was observed by inverted phase contrast microscope, and cell fusion and ciliated differentiation were observed by scanning electron microscope and immunocytochemistry. High-speed digital micro-video imaging system detects cilium swinging frequency. Results under the microscope, the number of ciliated cells increased gradually, and then decreased gradually after reaching the peak at day 7. The survival time of ciliated cells was maintained at 1421 days after 7 days of culture. Scanning electron microscopy showed that nasal epithelial cells were covered with ciliated or microvilli, goblet cells and non-ciliated columnar epithelial cells were arranged on the 7th day of culture, and type 鈪，

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