大鼠椭圆囊再生毛细胞前体细胞来源的初步研究

The Origination of Progenitor Cells of Utricular Regenerative Hair Cells in Rat

刘俊;孔维佳;赵荣祥;

1:浙江省中医院耳鼻咽喉科

2:华中科技大学同济医学院附属协和医院耳鼻咽喉科

摘要
目的探讨哺乳动物椭圆囊再生毛细胞前体细胞的可能来源。方法取出生1天的大鼠的椭圆囊,经嗜热菌蛋白酶(thermolysin)处理,消化法进行原代培养。在倒置显微镜下观察椭圆囊感觉上皮细胞(utricular sensory epichelial cell,USEC)的形态、生长特征;透射电镜观察、免疫细胞化学法检测上皮细胞角蛋白18、波形蛋白等鉴定USEC上皮细胞来源。免疫细胞化学法、RT-PCR技术检测支持细胞标记物p27kiplmRNA及毛细胞的特征性标记物Brn3a、Calretinin及AchRa9、MyosinⅦa mRNA的表达。结果原代培养的USEC呈扁平、多角形、核大而圆的上皮细胞形态,细胞之间连接紧密,形成单层时呈"铺路石样"外观。可见由数百个USEC包绕液体而成的dome(穹窿样)结构,表达细胞角蛋白,不表达波形蛋白,微绒毛丰富,细胞间连接紧密,提示其上皮来源。原代培养的USEC表达支持细胞标记物p27kiplmRNA及毛细胞的特征性标志物Brn3a、Calretinin及AchRa9、MyosinⅦa mRNA,表明培养的USEC可能来源于支持细胞并具有毛细胞的特性。结论原代培养的USEC能产生毛细胞样细胞且表达支持细胞标记物,表明其来源为支持细胞。椭圆囊感觉上皮的支持细胞可能为毛细胞再生的前体细胞之一。
关键词
椭圆囊;感觉上皮;再生;毛细胞
基金项目(Foundation):
国家自然科学基金(39925035);; 国家杰出青年基金(39925035);; 卫生部临床学科重点项目(2001321)基金资助
作者
刘俊;孔维佳;赵荣祥;
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刘俊孔维佳赵荣祥