Hath1基因转染后大鼠耳蜗毛细胞前体细胞的扫描电镜观察.pdfVIP

申国耳鼻喃喃魏辙稠/慧。06年5月，篇罐，.5. ·温- Hath1基因转染眉大鼠耳蜗毛细胞前体细胞的扫描电镜观察 张蝶，孙建和，郭维，胡吟燕，‘程所强 (解放军总医院耳鼻咽喉研究所，北京 100853) [摘要]目的探讨Hath1基因对大鼠耳蜗大上皮峭 (greater epithelial ridge , GER) 细胞的诱导分化作 用。方法取出生脑第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，子含10 % 血清DMEM墙基中进行培拌。以腺病毒为载体对GER培 养物进行Hath1暴囚的转换，并对感染脂10天的标本世 行扫描电镜观棋。描踉扫描电镜观察显示G巴R细胞在 含10 %血清DMEM培幕中黑片状贴黯生长恃性，表现 为典盟的上皮样务角型外观。 Hath1感染脂10天的GER 细胞培挥物可见在细胞团的边缔出现了细丝状纤毛样 结构，而无病毒感染对照组未见到该结构。结论 GER 细胞作为毛细胞前体细胞可以提现体外服代培养，在 HathH基因置在达情况下，部分细胞的寝面可见不成熟 的纤毛样结构，据来Hath1幕剧也许可以请导离休培养 下的纯的电细胞前体细胞向毛细胞的初步分化。 {关键词]毛细胞;再生;显微镜检查，电子，扫描;纤 毛 SEM observation of hair cell progenitors forced to express Hath1 from rat cochleae ZHANG γuan， SUN Jianhe, GUO Wei, HU Yinyan,ZHAI Suoqiang (Institute of Otolaryngology, Chinese PLA General Hospital, Beijing ,100853, China) Corresponding author: ZHAI Suoqiang(Email: 拍创sq@ ) [ABSTRACT] OBJECTIVE To investigate the e仔ect of Hath1 on epithelial ridge (GER) cells from postnatal rat cochlear. METHODS An experimen阳I method was developed which allowed the isolation and culture of GER cells from P1 rat cochleae using a combinatorial approach of enzymatic digestion and mechanical separation. The dissociated GER cells were cultured in DMEM+10% FBS. The GER explants infected with ad-Hath1 for 3 hours were cultured for 10 days and observed under scanning electron microscope. R应SULTS The GER cell cultures tended to attach to the substratum and grow in patches which assume a polygonal morphology similar to that of epithelial cells in medium containing 10% serum. Stereociliary bundle嗣 like structures were observed in the boundary of GER cell patches forced to express Hath1. CONCιUSION 丁hat GER cells which are likely hair cell progenitors could be cultured in vitro and generate stereociliary bundle.….Iike structure when forced to express Hath1 suggests that the misexpression of Hath1 probably can induce the predi忏erentiatíon of pure hair cell progenitors into hair cells in vitro. [Key words] Hair Cells;Regeneration;Microscopy,E