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After 24 h in fresh media, zeocin (Invitrogen) was added to the medium of cells for selection (100200 g/mL)

Aging causes changes in the cochlear tissue structure, including thickening of the basilar membrane, weakening of the elasticity of the spiral ligaments, and narrowing and hardening of the blood vessels, leading to impaired sound conduction, ischemia, and hypoxia in the cochlea

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To determine the kinetic constants, the enzyme concentration and concentrations of its substrates were first optimized so that the absorbance and fluorescence intensity remained linear