免疫磁珠纯化人鼻黏膜上皮细胞的实验研究  

Purification of human nasal epithelial cells by using immunomagnetic beads

作　　者：陈振振[1] 贡欣[2] 陆洋[1] 杜守颖[1] 蔡程博[1] 尚可心[1] 

机构地区：[1]北京中医药大学中药学院,北京100102 [2]北京中医药大学东方医院妇科,北京100078

出　　处：《重庆医学》2014年第20期2609-2612,共4页Chongqing medicine

基　　金：国家自然科学基金项目(81073057);教育部博士点基金(20090013110007);北京中医药大学创新团队项目(2011-CXTD-13)

摘　　要：目的建立高纯度人鼻黏膜上皮细胞(HNEC)的体外培养方法,为鼻腔制剂的评价提供良好的细胞模型。方法采用酶消化法分离细胞、进行无血清培养。磁珠分选法选择HNEC群,流式细胞仪鉴定纯化前后上皮细胞纯度,扫描电镜下进行形态学观察。结果分离培养的细胞光镜下呈铺路鹅卵石状,电镜下可见丰富的微绒毛和纤毛分化,经流式细胞仪检测鉴定分选后所得细胞纯度达到99%以上。结论磁性分选系统可以直接分离得到高纯度的HNEC,该模型可为鼻腔制剂的药物基础性研究提供实验基础。Objective To establish a high purity primary culture methods of human nasal epithelial cells （HNEC） in vitro and to provide a successful primary culture model for evaluation experiments of the nasal preparation .Methods Primary culture of human nasal epithelial cells were performed with enzymatic dissociation of isolated tissue and cultured in serum-free medium .HNEC were separated through magnetic field by immunomagnetic beads .We determined the purity of the separated cells by light microscopy and flow cytometry .The morphology of HNEC was observed with a scanning electron microscope .Results Under an inverted phase microscope ,the cells morphology was paving stone shaped .Under the scanning electron microscopy ,abundant microvilli and cilia differentiation were observed .Flow cytometry showed the epithelial cells accounted for 99% .Conclusion The highly purified HNEC can be directly isolated by the magnetic cell sorting system .The cell model can be used for the basic research of nasal cavity preparation .

关 键 词：鼻黏膜 上皮细胞 原代培养 免疫磁珠 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]