新生小鼠耳蜗Corti器的体外培养与观察.pdfVIP

DOC格式论文，方便您的复制修改删减 新生小鼠耳蜗Corti器的体外培养与观 察 （作者:___________单位: ___________邮编: ___________） 作者：许涛,孟秀清，王爱梅,刘双月,牟华 【摘要】目的建立新生昆明小鼠耳蜗 Corti 器体外培养的简便方 法并观察耳蜗毛细胞的组织学形态。方法 取生后3 ～5 d 昆明小鼠耳 蜗基底膜，平铺在胶原凝胶表面，然后在含有 1% 牛血清白蛋白的 DMEM/F12 培养基中进行培养。采用异硫氰酸四甲基罗丹明标记的 鬼笔环肽荧光染色方法观察耳蜗毛细胞的生长情况。结果 耳蜗基底 膜经24、48 h 和72 h 培养后，内、外毛细胞均生长良好，其结构完 整，纤毛束排列有序，无任何缺失。结论 本培养方法简便有效，可 用于耳蜗Corti 器体外培养实验研究。 【关键词】小鼠;耳蜗;体外培养;Corti 器 Abstract: Objective To establish an simple method for the culture of the organ of Corti from newborn Kunming mouse, and to observe the histological morphology of cochlear hair cells. Methods The cochleae from Kunmin mice of postnatal day 3 to 5 were used. DOC格式论文，方便您的复制修改删减 The basilar membrane of the mouse was isolated and positioned on the surface of collagen gel, and then cultured in DMEM / F12 medium containing 1% bovine serum albumin. Tetramethylrhodamine isothiocyanate-labeled phalloidine staining was used to observe growing status of hair cells. Results Following culture of the cochlear basilar membrane for 24 hours, 48 hours and 72 hours, the inner and outer hair cells were both growing well. Their structures were intact, and stereociliary bundles were arranged orderly, without any deletion. Conclusions This method is simple and effective, which can be applied to the experimental study of the organ of Corti in vitro culture. Key words:mouse; cochlea; culture; organ of Corti 耳蜗 Corti 器体外培养是进行内耳生理和病理生理等实验研究 的重要手段，尤其对研究各种耳毒性药物的听觉损伤机制及其防治办 法具有较大的实际应用价值。目前关于耳蜗Corti 器体外培养的方法 各不相同，本文介绍一种简便有效的新生昆明小鼠耳蜗Corti 器的体 外培养方法，并通过异硫氰酸四甲基罗丹明(Tetramethylrhodamine isothiocyanate ，TRITC)标记的鬼笔环肽荧光染色技术，观察了耳蜗 毛细胞的组织学形态。 1 材料与方法 1.1 实验动物 选择出生3 ～5 d 的昆明小鼠20 只，雌雄不限，由辽宁医学院 DOC格式论文，方便您的复制修改删减 实验动物中心提供。 1.