[Inner hair cells isolation and morphological observation].
OBJECTIVE
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.


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 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.


CONCLUSIONS
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.
Yang
Shiming
S
Department of Otorhinolaryngology Head and Neck Surgry, Institute of Otolaryngology, PLA General Hospital, Beijing 100853, China. shim@public3.bta.net.cn
Shen
Jing
J
Yamashita
Toshio
T
Yang
Weiyan
W
Gu
Rui
R
Han
Dongyi
D
D000818:Animals; D002469:Cell Separation / Q000379:methods; D003051:Cochlea / Q000166:cytology*; D005260:Female; D006168:Guinea Pigs; D006199:Hair Cells, Auditory, Inner / Q000166:cytology*; D018072:Hair Cells, Auditory, Outer / Q000166:cytology; D008297:Male