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目的 在离体状态下对豚鼠前庭椭圆囊器官进行培养,观察庆大霉素损伤毛细胞后细胞的增殖情况。方法采用白色健康豚鼠36只,体重300-450克,随机分为4组:(1)正常对照组;(2)正常加BrdU(5-溴基脱氧尿核苷BrdU)组;(3)庆大霉素组;(4)庆大霉素加BrdU组。应用体外组织培养、5—溴基脱氧尿核苷(BrdU,3μg/ml)掺入及免疫组织化学染色和Epon树酯包埋半薄切片等方法,在不同时间(2、3、5、15天)光镜观察庆大霉素(2.0mM)损伤后毛细胞及支持细胞增殖情况。结果 用庆大霉素48小时后,可见到椭圆囊感觉上皮中的毛细胞溶解破坏。加入BrdU继续培养,BrdU免疫组织化学染色显示了第5和15天被BrdU标记的阳性细胞核,阳性细胞核多位于椭圆囊感觉上皮中基底层,从细胞的形态和分布位置上看为支持细胞;感觉上皮的表层,也可观察到BrdU反应阳性的上皮细胞,细胞核较圆,形态和位置类似毛细胞。结论 提示损伤后的椭圆囊感觉上皮支持细胞可以进行细胞的有丝分裂,支持细胞可能是修复毛细胞的前体细胞。
OBJECTIVE To culture the guinea pig vestibular elliptic sac organ in vitro and observe the proliferation of gentamicin-damaged hair cells. Methods 36 white healthy guinea pigs weighing 300-450 g were randomly divided into 4 groups: (1) normal control group; (2) BrdU group; (3) BrdU group Gentamicin group; (4) gentamicin plus BrdU group. In vitro tissue culture, incorporation of 5-Bromodeoxyuridine (BrdU, 3μg / ml) and immunohistochemical staining and Epon resin embedding semi-thin sections and other methods at different times (2,3,5,15 Day) light microscope to observe the proliferation of hair cells and supporting cells after gentamicin (2.0mM) injury. Results 48 hours after gentamicin, can be seen in the epithelial oval capsule hair cell destruction. After adding BrdU for further culture, BrdU immunohistochemical staining showed positive BrdU-labeled nuclei on the 5th and 15th days. The positive nuclei were mostly located in the basal layer of the epicardial sensory epithelium of the oval sac and were seen as supporting cells from the morphological and distributional locations of the cells. Epidermal epithelium, but also can be observed in BrdU positive epithelial cells, the nucleus is round, the shape and location similar to hair cells. Conclusions It is suggested that the epithelial cells in the oval sac after injury may be mitotic cells, and the supporting cells may be the precursors to repair the hair cells.