中华耳科学杂志Issue(1)：76-78,3.DOI:10.3969/j.issn.1672-2922.2015.01.013
PET滤膜用于承载耳蜗基底膜培养的实验研究
Polyethylene Terephthalate Membrane Used as a Holder for Cochlear Organotypic Cultures
摘要
Abstract
Objective To report a practical method for tissue adhere in cochlear organotypic cultures. Methods The polyethylene terephthalate (PET) membrane was precoated with laminin, poly-L-ornithine, and fetal bovine serum at a ratio of 2:1:1 overnight. The cochlear basilar membrane was micro-dissected from C57BL/6J mouse pups at postnatal day 4, and placed on the PET membrane in a culture medium containing DMEM, 10%fetal bovine serum, and 30U/ml penicillin. After 48 hours, the cochlear specimens were fixed with 10%formalin in PBS. Anti-otoancorin and anti-tubulinⅢantibodies were dou⁃ble stained for labeling of nonsensory cells, interdental cells in the inner ear and auditory nerve fibers respectively. Fluores⁃cence conjugated phalloidin was also used to label stereocilia and cuticular plate of hair cells, and DAPI was also applied for nuclear staining. In addition, cochlear explants were also examined using scanning and transmission electron microscopy. Re⁃sults The cochlear basilar membrane firmly attached to the surface of PET. The microstructure and ultrastructure of cochlear cells appeared normal. Conclusion PET membrane can be used as an ideal holder for cochlear organotypic cultures.关键词
聚对苯二甲酸乙二醇酯(PET)滤膜/耳蜗器官培养/毛细胞/齿间细胞/免疫荧光/透射电镜/扫描电镜Key words
Polyethylene terephthalate membrane/Cochlear organotypic cultures/Hair cell/Interdental cell/Immunocy-tochemistry/Scanning electron microscopy/Transmission electron microscopy分类
医药卫生引用本文复制引用
刘永泽,周函,麻晓峰,马登滨,高下..PET滤膜用于承载耳蜗基底膜培养的实验研究[J].中华耳科学杂志,2015,(1):76-78,3.基金项目
江苏省自然科学基金资助(项目号BK2010104) （项目号BK2010104）
