内毛细胞的分离和形态学特点  

Inner hair cells isolation and morphological observation

作　　者：杨仕明[1] 沈静[2] 山下敏夫[3] 杨伟炎[1] 顾瑞[1] 韩东一[1] 

机构地区：[1]解放军总医院耳鼻咽喉头颈外科解放军耳鼻咽喉科研究所,北京100853 [2]中国医科大学听力研究室 [3]日本关西医科大学耳鼻咽喉科

出　　处：《中华耳鼻咽喉科杂志》2002年第6期462-464,I005,共4页Chinese Journal of Otorhinolaryngology

基　　金：国家自然科学基金 ( 30 0 0 0 189);军队"十五"科研基金( 0 1Q0 5 0 );解放军总医院"中青年学科带头人工程"课题基金资助

摘　　要：目的　探讨内毛细胞 (innerhaircells ,IHC)的分离技术和形态学特征。方法　从豚鼠耳蜗制备基底膜活体铺片 ,微分干涉倒置显微镜下用显微分离结合酶消化的方法单离IHC。高倍显微镜下 ,用两个钨丝电极沿IHC和第一排外毛细胞 (outerhaircells,OHC)之间的Corti隧道显微切割 ,游离的IHC团被吸引至玻璃微吸管内。结果　从每只豚鼠耳蜗可分离到 3 0～ 5 0个活性良好的单离IHC ,并存活 2h左右。典型的IHC胞体具有特征性 ,呈梨状或长颈的烧瓶形状 ,胞体中间可见大的圆形细胞核。胞体内包含较多细胞器 ,颗粒状 ,粗糙感。大部分可观察到静纤毛 ( 93 / 98)。静纤毛分布沿表皮板一侧 ,排列呈线状或“C”形。部分细胞可观察到明显狭窄的颈部 ( 61/ 98)。IHC的表皮板一般为倾斜状 ,凹面向上 ,较厚。另外 ,IHC的表皮板通常与细胞胞体长轴成锐角或钝角。豚鼠耳蜗IHC长度为 13～ 3 1μm ,平均 ( 2 2 45± 4 14) μm ( x±s,n =98,下同 ) ;直径为 7～ 15 μm ,平均 ( 11 95±1 5 9) μm。静纤毛长度为 2～ 7 5 μm ,平均 ( 5 2 1± 1 0 0 ) μm。 结论　本研究成功建立一种能够分离到大量的、生物活性好的IHC的技术 ;内、外毛细胞单离后在形态学上的鉴别要点是表皮板 。Objective To set up an effective technique for isolating the single inner hair cells (IHCs) and observe morphological features to distinguish IHCs from outer hair cells (OHCs) in vitro. Methods Surface preparations contained rows of OHCs and IHCs were prepared from guinea pigs. The cluster and single IHCs were separated microsurgically combination with an enzymatic digestion. The clusters of IHCs were separated using two fine electrodes and transferred by a gentle suction using a glass micropipette. Results On the average, one guinea pig cochlear yielded approximately 30 to 50 viable solitary IHCs. The cell body of the IHCs was pear or flask shape. The nucleus was located in the central of the cell body, and the cytoplasm was filled with the structured and scattered rough granules. The stereocilia were seen in the most of IHCs (93 out of 98). The stereocilia were aligned, almost lineally or in a 'C' shape, in one side on the surface of the cuticular plate. The tight neck and the angle between the cuticular plate and the axis of the cell were observed in some IHCs but not obvious in most cases, which might be due to the orientation of the cells. The length of isolated IHCs was ranged from 13 to 31 μm and with an average of (22.45±4.14)μm (±s,n=98). The diameter of IHCs was ranged from 7 to 15 μm and with an average of (11.95±1.59)μm. The length of stereocilia was ranged from 2 to 7.5 μm and with an average of (5.21±1.00)μm. Conclusion Unambiguous solitary IHCs were successfully harvested from guinea pig cochlea using a microsurgical technique. The tight neck and the angle between the cuticular plate and the axis of the IHCs have been considered as an important landmark to distinguish the IHCs from OHCs.

关 键 词：内毛细胞 分离 形态学 

分 类 号：R764[医药卫生—耳鼻咽喉科]