豚鼠内耳椭圆囊斑体外原代培养

Primary organ culture of the utricular maculae of the guinea pig inner ear in vitro

目的探讨豚鼠内耳椭圆囊斑体外正常培养状态及其在体外用氨基糖苷类抗生素庆大霉素后的毒性损伤特征,以期建立前庭外周器官体外实验模型。方法豚鼠被随机分成三组:①正常未培养对照组;②正常原代培养组,利用体外组织培养方法对豚鼠内耳椭圆囊斑进行正常原代培养;③庆大霉素损伤原代培养组。正常原代培养组和庆大霉素损伤原代培养组在培养期间每天用相差显微镜观察生长晕,正常培养组在培养的第5天和第10天固定用石蜡包埋切片光镜观察。庆大霉素组在48小时用树脂包埋半薄切片观察。结果正常原代培养组在培养10天过程中,用相差显微镜观察椭圆囊斑在培养第1～4天无明显生长晕,在培养第5天明显从椭圆囊边缘长出。随培养时间延长,生长晕不断扩大,以成纤维细胞为主。庆大霉素损伤原代培养组在培养2天内每天用相差显微镜观察椭圆囊斑未见明显生长晕。正常原代培养至第5和第10天,石蜡包埋切片显示支持细胞和毛细胞存活。庆大霉素组在48小时,用树酯包埋半薄切片观察,感觉上皮层的毛细胞溶解破坏,支持细胞存活。结论椭圆囊斑器官体外培养方法可行,并培养成功,为今后进一步研究椭圆囊斑提供了实验模型。

OBJECTIVE To explore condition of primary organ culture of the utricular maculae of the guinea pigs inner ear in vitro and establish an experimental model for further research. METHODS The utricular maculae of the guinea pigs were primarily cultured with tissue culture method in vitro. RESULTS The grow of the outgrowth zone of the utricular maculae presented fibroblasts cell during 10 days, Existing of the supporting cells and hair cells in sensory epithelium were found by means of paraffin embedded sections on 10th day. Hair cells were damaged and supporting cells were lived in culture medium with gentamicin for 48 hours by means of plastic embedded mount and semi-thin sectioning, CONCLUSION Primary organ culture of the utricular maculae of the guinea pig inner ear in vitro is feasible, This organ culture method provide an experimental model for further research on utricular maculae,