The potential use of stem cells to repair hearing loss requires surgical access to the cochlea. Here we describe a microsurgical technique for cell injection into the mouse cochlea. Green fluorescent cells (ZsGreen-MCF10A cells) were successfully injected via a lateral wall cochleostomy into the scala media, scala tympani and scala vestibuli compartments of the cochlea. The effect of surgery on auditory function was investigated with auditory brainstem responses (ABR) to click and tone stimuli. A computerised signal-to-noise ratio detection method was developed to measure ABR thresholds in conjunction with visual inspection. Signal-to-noise ratio detection showed ABR thresholds in control mice were similar for click (33+/-7 dB) and tone stimuli (33+/-6 dB), in agreement with visual inspection (click 39+/-7 dB, tone 35+/-6 dB). The mean ABR threshold for combined click and tone stimuli was 15-45 dB greater after surgery with minimum hearing loss achieved with a small sized cochleostomy (< or =0.4mm) andby sibling matching to control mice (control 33+/-4 dB, surgery 48+/-3 dB). The microsurgical technique will provide a basis for future studies on the use of stem cells in the treatment of hearing loss.
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