Hath1基因诱导新生大鼠大上皮嵴细胞形成毛细胞样细胞  

Overexpression of Hathl induces production of hair cell-like cells in greater epithelial ridge cell cultures from postnatal rat cochlea

作　　者：张媛[1] 胡吟燕[1] 郭维[1] 翟所强[1] 

机构地区：[1]解放军总医院耳鼻咽喉科研究所,北京100853

出　　处：《中华耳鼻咽喉头颈外科杂志》2008年第5期360-363,共4页Chinese Journal of Otorhinolaryngology Head and Neck Surgery

基　　金：国家自然科学基金（30440063）;海外青年学者合作研究基金（30228029）;北京市自然科学基金（7053076）

摘　　要：目的探讨Hathl（human atonal homolog 1）基因对大鼠耳蜗大上皮嵴（greater epithelial ridge，GER）细胞的诱导分化作用。方法取出生后1d大鼠耳蜗，利用酶消化和机械分离相结合的方法分离出GER细胞，并行体外培养。以腺病毒为载体，对GER细胞培养物进行Hath1基因和增强型绿色荧光蛋白（enhanced green fluorescent protein，EGFP）基因（ad—Hath1-EGFP）的转染，单纯转染EGFP（ad-EGFP）作为对照组，对感染后不同天数的标本行毛细胞特异性标记物myosin Ⅶa免疫组化染色鉴定。结果ad-Hath1—EGFP组中的GER细胞中出现了myosin Ⅶa和EGFP双标记细胞，并且从感染后第5天开始出现myosin Ⅶa阳性细胞，随后阳性细胞的数量有所增加，但增加不明显；而adEGFP组感染后3～12d的GER标本均未见myosinWa阳性细胞。结论GER细胞可能是耳蜗毛细胞的前体细胞，Hath1基因过表达可以诱导GER细胞向毛细胞样细胞（myosin Ⅶa阳性）分化。Objective To investigate the effect of Hathl （human atonal homolog 1） overexpression on greater epithelial ridge （GER） cells from postnatal rat cochlea in vitro. Methods GER cells were isolated by using a combinatorial approach of enzymatic digestion and mechanical separation from P1 rat cochlear. The GER cell cultures were infected by adenovirus containing Hathl and enhanced green fluorescent protein （ad-Hath1-EGFP）, while transfecting EGFP（ad-EGFP）was as controls. Immunostaining were performed at different time points after adenovirus infection. Results Some of the infected GER cells became myosin Ⅶa-positive following ad-Hath1-EGFP infection. The earliest time point to see induction of hair cell differentiation （ hair cell marker expression） by ad-Hath1 was 5 days post-infection. In contrast, infection of the GER sheet cultures with ad-EGFP control virus did not show any myosin Ⅶa-positive cells at 3 - 12 days post-infection in all cultures examined. Conclusions GER cells may potentially serve as hair cell progenitors and they are capable of differentiating hair cell-like cells when forced to express Hathl.

关 键 词：耳蜗 细胞 培养的 转染 Hath1 

分 类 号：R686[医药卫生—骨科学]