Impairment Effect of Kanamycin on Mouse Cochlear Hair Cells in Vitro

Objective To establish an organ culture model of Corti to investigate the impairment effect of kanamycin(KM)on mouse cochlear hair cells.Methods The cochlear basilar membrane from 3-day-old mouse were isolated and positioned on the surface of collagen gel and cultured in DMEM/F12 medium containing 1% bovine serum albumin.Tetramethylrhodamine isothiocyanate-labeled phalloidine staining was used to observe the effects of different concentration of KM(0.05,0.1,0.2,0.3 mmol/L)on mouse cochlear hair cells.The loss of hair cells was detected by cell counts.Results The cilia of cochlear outer hair cells were lodged and arranged in disorder caused by different concentration of KM.When KM concentration of was 0.05 mmol/L,the percent of missing outer hair cell was significantly increased as compared with control group(P0.05),while the percent of missing inner hair cell was no significant changed(P0.05).With KM concentration was gradually increased(0.1~0.3 mmol/L),the percents of missing inner and outer hair cell were significantly increased,showing a clear dose-esponse relationship(P0.05 or P0.01).Conclusion KM could impair hair cells in mouse cochlear culture,and organ culture of inner ear is a effective method for studying drug ototoxicity.