转基因诱导新生大鼠耳蜗大、小上皮嵴细胞分化为毛细胞样细胞实验  

Differentiation of newborn rat cochlear greater and lesser epithelial ridge cells into hair cell-like cells induced by gene transfection

作　　者：翟所强[1] 张媛[2] 汪学勇[3] 宋巍 郑贵亮[5] 胡吟燕[1] 郭维[1] 孙建和[1] 

机构地区：[1]解放军总医院耳鼻咽喉头颈外科医院研究所,北京100853 [2]北京同仁医院耳鼻咽喉头颈外科,北京100730 [3]北京世纪坛医院耳鼻咽喉科,北京100038 [4]徐州市人民医院耳鼻咽喉科,徐州221003 [5]上海长海医院耳鼻咽喉科,上海200433

出　　处：《中华耳科学杂志》2009年第1期1-4,共4页Chinese Journal of Otology

基　　金：海外青年学者合作基金(30228029);十一五国家科技支撑课题(2006BAI02B06);国家自然科学基金(30772413)资助

摘　　要：目的通过转染Hath1基因诱导大鼠耳蜗大上皮嵴细胞(greater epithelial ridge,GER)和小上皮嵴细胞(lesser epithelial ridge,LER)分化为毛细胞样细胞。方法取出生后第1天的大鼠耳蜗,利用机械分离和酶消化相结合的方法分别分离出纯的GER和LER细胞,并转染Hath1基因,培养后做免疫组化染色观察。结果GER和LER体外培养并转染Hath1基因后,免疫组化检测显示肌球蛋白(myosin Ⅶa)染色阳性,表达毛细胞的特异标记物,提示GER和LER已分化为毛细胞样细胞。结论GER和LER细胞作为毛细胞前体细胞可以实现体外培养,转染Hath1后,可分化为毛细胞样细胞。Objective To study differentiation of cells from the greater epithelial ridge (GER) and lesser epithelial ridge (LER) into hair cell-like cells. Methods Using a combinatorial approach of enzymatic digestion and mechanical separation, GER and LER cells from P1 rat cochleae were isolated. The dissociated GER and LER cells were infected with Hath1 and cultured. The infected cells were examined using immunohistochemistry. Results The GER and LER cells clearly showed capability to proliferate. When infected with Hath1, the GER and LER cells stained positive for Myosin VIIa, suggesting the potential to differentiate into hair cell-like cells. Conclusion The GER and LER cells are likely hair cell progenitors and can differentiate into hair cell-like cells when infected with Hath1.

关 键 词：毛细胞 再生 大上皮嵴 小上皮嵴 

分 类 号：R322.923[医药卫生—人体解剖和组织胚胎学] R329.29[医药卫生—基础医学]