建立大鼠前庭器官体外培养模型及观察佛波酯作用下的超微结构变化

Establishment of rat vestibular organ culture system in vitro and observation of ultrastructure change induced by phorbol myristate acetate

目的：建立大鼠前庭器官的原代体外培养模型，并通过透射电镜观察前庭上皮细胞在佛波酯（PMA）作用前后的超微结构变化．方法：无菌条件下取出生2d大鼠的椭圆囊上皮，体外培养1～7d；从第2日开始用倒置显微镜观察、照相，试验组加入PMA30min，2h和6h，固定，透射电镜观察．结果：体外培养椭圆囊1wk，12h时可见成纤维细胞沿移植物边缘游出，2d后内皮细胞聚集，呈铺路石样，透射电镜观察细胞无肿胀，胞膜、胞核完整，线粒体嵴无肿胀，纤毛无脱落；PMA各试验组细胞形态正常，核糖体密集，高尔基体较前发达，内质网轻度扩张．结论：大鼠前庭器官体外培养方法可行，为今后进一步研究提供了实验模型．

AIM： To establish a culture system of rat vestibular organ in vitro and observe the change of cell structure after phorbol myristate acetate （PMA） induction by transmission electron mi croscope. METHODS： The vestibular epithelia of postnatal 2day rats were isolated and cultured in Dulbecco＇s modified Eagle＇s medium and observed by inverted microscope. After induced by PMA, the cultured cells were all assessed by transmission electron microscopy. RESULTS： The vestibular organ exhibited a “cobblestone-like” appearance after 2 d , while the ultrastructure of culture group was normal after 7 d . In the PMA-induced group, the tested cells had normal shape, but the changes of ribosome, Golgi body and endoplasmic reticulum were observed. CONCLUSION： The change of ultrastructure of PMA-induced group has a relationship with the expression level of cellular protein kinase. The establishment of rat vestibular organ culture model in vitro lays a foundation for the further study of inner ear disease.