大鼠内耳前庭上皮细胞的体外培养研究
In vitro culturing of inner ear vestibular epithelial cells of SD rats
马瑜徽 1徐丽华 1周鑫 1王国华 1姜正林1
作者信息
- 1. 南通大学特种医学研究院,南通 226019
- 折叠
摘要
目的 建立大鼠内耳前庭上皮细胞的体外培养体系,为进一步研究内耳前庭相关疾病及其机制奠定基础.方法 实验选用7 d龄SD大鼠乳鼠,活体分离出球囊和椭圆囊组织,消化处理破碎后产生的细小组织块放置在细胞培养箱中进行培养.体外培养5～7d后,采用免疫荧光法检测上皮细胞标志物角蛋白7(CK7)和支持细胞标志物Lgr5的表达及纯度.结果 体外培养的前庭上皮细胞呈不规则多角形,紧密相连呈团状,周围有少量成纤维细胞.绝大多数细胞可成功标志CK7和Lgr5,且主要在胞质中表达.成功标记CK7和Lgr5的细胞阳性率均在90％以上,上皮细胞纯度较高.结论 采用组织块培养技术,在体外建立前庭上皮细胞培养体系,为进一步研究内耳前庭功能、内耳疾病的病理生理机制和毛细胞再生提供合适的细胞体系.
Abstract
Objective To establish an in vitro culture system of inner ear vestibular epithelial cells(ECs)of Sprague-Dawley(SD)rats,so as to lay a foundation for further research on inner ear vestibular system-related diseases and their mechanisms.Methods For the experiment,7-day-old SD suckling rats were selected,and the saccule and utricle tissues were isolated from the living rats.These tissues were digested and fragmented into small pieces,which were then cultured in a cell incubator.After 5-7 days of in vitro culturing,the expression and purity of epithelial cell marker cytokeratin 7(CK7)and Sertoli cell marker Lgr5 were detected by using immunofluorescence.Results ECs cultured in vitro presented irregular polygonal shape,being closely connected in clusters,and surrounded by a small quantity of fibroblasts.The majority of cells were successfully labeled with CK7 and Lgr5,with expression primarily observed in the cytoplasm.The positive rate of cells successfully labeled with CK7 and Lgr5 was over 90％,indicating a high purity of epithelial cells.Conclusion The vestibular epithelial cell culture system was established in vitro through tissue-block culture technology,thereby providing a suitable cell model for further research on the vestibular functions and pathophysiological mechanisms of inner ear diseases and hair cell regeneration.
关键词
上皮细胞/前庭/内耳/细胞培养Key words
Epithelial cell/Vestibule/Innerear/Cell culture引用本文复制引用
出版年
2024
NETL
NSTL
万方数据