诱导新生大鼠耳蜗大上皮嵴细胞分化为毛细胞样细胞  

Induced differentiation into hair cell-like cells from the greater epithelial ridge of the newborn rat cochlea

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

机构地区：[1]解放军总医院耳鼻咽喉头颈外科研究所,北京100853 [2]首都医科大学附属北京同仁医院耳鼻咽喉头颈外科,耳鼻咽喉头颈科学教育部重点实验室(首都医科大学),北京100730

出　　处：《中国耳鼻咽喉头颈外科》2009年第11期599-603,共5页Chinese Archives of Otolaryngology-Head and Neck Surgery

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)联合资助

摘　　要：目的大鼠耳蜗大上皮嵴细胞(greater epithelial ridge,GER)转染Hath1基因,与耳蜗间质细胞共培养诱导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离出纯的GER细胞,与耳蜗间质细胞共培养或转染Hath1基因培养,做免疫组化和扫描电镜观察。结果GER体外培养具有增殖能力,GER与耳蜗间质细胞共培养或转染Hath1基因后,免疫组化MyosinVIIa阳性,扫描电镜部分细胞的表面可见不成熟的纤毛样结构,提示GER已分化为毛细胞样细胞。结论GER细胞作为毛细胞前体细胞可以实现体外培养,与耳蜗间质细胞共培养或转染Hath1后,可分化为毛细胞样细胞。OBJECTIVE To transfect Hath1 gene to greater epithelial ridge（GER）cells, and co-cultured with mesenchymal cells for inducing them into hair celllike cells. METHODS GER cells were isolated and cultured from P1 rat cochlea using enzymatic digestion and mechanical separation methods. The dissociated GER cells were co-cultured with mesenchymal cells and transfected with Hath1 gene, observed under scanning electron microscope, and studied with immunohistochemistry method. RESULTS The GER cells presented ability of proliferation. Myosin VIIa was positive in GER cells, when they were co-cultured with mesenchymal cells or transfected with Hath1 gene. Stereociliary bundle-like structures were observed in the boundary of GER cells. That showed they had the potential to differentiate into hair cell- like cells CONCLUSION The GER cells, as progenitors of hair cells, can be cultured in vitro. When co-cultured with mesenchymal cells or transfected with Hath1 gene, GER cells can be induced differentiation into hair celllike cells.

关 键 词：大鼠 动物 实验 毛细胞 再生 显微镜检查 电子 扫描 

分 类 号：R764[医药卫生—耳鼻咽喉科]