摘要
目的：建立大鼠耳蜗大、小上皮嵴细胞永生细胞系。方法：取P1大鼠耳蜗，分离出耳蜗上皮薄片组织，并与含SV40-LT抗原的逆转录病毒共培养，从而实现细胞的永生化。利用单克隆法纯化细胞系，并通过形态学、免疫组织化学等方法对细胞系进行鉴定。结果：大鼠耳蜗大、小上皮嵴细胞永生细胞系基本保持了细胞原代的表型特征：呈现上皮细胞样的多角形外观；不同世代的细胞均表达Isletl及SV40-LT抗原。该系目前已培养3个月，超过20代，传代时间为3～4d，传代比例为1：5。结论：本实验成功建立了大鼠耳蜗大、小上皮嵴细胞的永生细胞系，这为大、小上皮嵴细胞生物学特性及相关功能，特别是毛细胞再生及前体细胞移植治疗感音神经性聋方面的研究奠定了坚实的实验基础。
Objective：To facilitate further investigation of the function of greater epithelial ridge （GER） and lesser epithelial ridge （LER） cells, a greater a lesser epithelial ridge cell immortal line originated from postnatal rat cochleae were established. Method：The GER and LER cells were dissected from P1 rats cochleae and infected with SV40-LT antigen using a retrovirus-mediated method. The established cell lines were purified by monoclone technique and identified following morphology observation and immunnocytochemical staining. Result： The established cell lines had been stably maintained continuously for more than 20 passages and displayed many features similar to epithelial cells. They grew in patches and assumed a polygonal morphology. Immunocytochemical characterization revealed that these cells could be labeled by GER/LER marker （Isletl） and SV40-LT antigen. Conclusion： In the present experiment, the GER and LER cell line from postnatal rat cochleae has been successfully established and this cell line may potentially provide an invaluable system for studying hair cell differentiation and regeneration as well as the therapy of sensory deafness caused by auditory hair cells loss.
出处
《临床耳鼻咽喉科杂志》
CSCD
北大核心
2006年第10期460-462,465,共4页
Journal of Clinical Otorhinolaryngology
基金
海外青年学者合作研究基金资助项目(No:30228029)
国家自然科学基金资助项目(No:30440063)
北京市自然科学基金资助项目(No:7053076)
关键词
耳蜗
大上皮嵴
小上皮嵴
永生细胞系
Cochlea
Greater epithelial ridge
Lesser epithelial ridge
Immortalized cell lines
