Yang Shiming, Shen Jing, Yamashita Toshio, Yang Weiyan, Gu Rui, Han Dongyi
Department of Otorhinolaryngology Head and Neck Surgry, Institute of Otolaryngology, PLA General Hospital, Beijing 100853, China.
Zhonghua Er Bi Yan Hou Ke Za Zhi. 2002 Dec;37(6):462-4.
To set up an effective technique for isolating the single inner hair cells (IHCs) and observe morphological features to distinguish IHCs from other hair cells (OHCs) in vitro.
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.
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 microns and with an average of (22.45 +/- 4.14) micron (mean +/- s, n = 98). The diameter of IHCs was ranged from 7 to 15 microns and with an average of (11.95 +/- 1.59) micron. The length of stereocilia was ranged from 2 to 7.5 microns and with an average of (5.21 +/- 1.00) micron.
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.
建立一种有效的分离单个内毛细胞（IHC）的技术，并观察其形态特征以在体外将IHC与其他毛细胞（OHC）区分开来。
从豚鼠制备包含成排OHC和IHC的表面标本。通过显微手术结合酶消化分离细胞簇和单个IHC。使用两个精细电极分离IHC细胞簇，并使用玻璃微吸管通过轻柔抽吸进行转移。
平均而言，一只豚鼠耳蜗可产生约30至50个存活的单个IHC。IHC的细胞体呈梨形或烧瓶形。细胞核位于细胞体中央，细胞质中充满了有结构的和分散的粗颗粒。在大多数IHC（98个中的93个）中可见静纤毛。静纤毛在角质板表面的一侧排列，几乎呈线性或呈“C”形。在一些IHC中观察到紧密的颈部以及角质板与细胞轴之间的角度，但在大多数情况下不明显，这可能是由于细胞的取向所致。分离的IHC长度范围为13至31微米，平均为（22.45±4.14）微米（平均值±标准差，n = 98）。IHC的直径范围为7至15微米，平均为（11.95±1.59）微米。静纤毛的长度范围为2至7.5微米，平均为（5.21±1.00）微米。
使用显微手术技术成功地从豚鼠耳蜗中收获了明确的单个IHC。紧密的颈部以及角质板与IHC轴之间的角度被认为是将IHC与OHC区分开来的重要标志。
