Hair cell regeneration is the fundamental method of correcting hearing loss and balance disorders caused by hair cell damage or loss. How to promote hair cell regeneration is a hot focus in current research. In mammals, cochlear hair cells cannot be regenerated and few vestibular hair cells can be renewed through spontaneous regeneration. However, Math1 gene transfer allows a few inner ear cells to be transformed into hair cells in vitro or in vivo. Hair cells can be renewed through two possible means in birds: supporting cell differentiation and transdifferentiation with or without cell division. Hair cell regeneration is strongly associated with cell proliferation. Therefore, this study explored the relationship between Math_1-induced vestibular hair cell regeneration and cell division in mammals. The mouse vestibule was isolated to harvest vestibular epithelial cells. Ad-Math_1-enhanced green fluorescent protein(EGFP) was used to track cell division during hair cell transformation. 5-Bromo-2′-deoxyuridine(Brd U) was added to track cell proliferation at various time points. Immunocytochemistry was utilized to determine cell differentiation and proliferation. Results demonstrated that when epithelial cells were in a higher proliferative stage, more of these cells differentiated into hair cells by Math_1 gene transfer. However, in the low proliferation stage, no Brd U-positive cells were seen after Math_1 gene transfer. Cell division always occurred before Math_1 transfection but not during or after Math_1 transfection, when cells were labeled with Brd U before and after Ad-Math_1-EGFP transfection. These results confirm that vestibular epithelial cells with high proliferative potential can differentiate into new hair cells by Math_1 gene transfer, but this process is independent of cell proliferation.
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