In vitro culturing of inner ear vestibular epithelial cells of SD rats
Objective To establish an in vitro culture system of inner ear vestibular epithelial cells(ECs)of Sprague-Dawley(SD)rats,so as to lay a foundation for further research on inner ear vestibular system-related diseases and their mechanisms.Methods For the experiment,7-day-old SD suckling rats were selected,and the saccule and utricle tissues were isolated from the living rats.These tissues were digested and fragmented into small pieces,which were then cultured in a cell incubator.After 5-7 days of in vitro culturing,the expression and purity of epithelial cell marker cytokeratin 7(CK7)and Sertoli cell marker Lgr5 were detected by using immunofluorescence.Results ECs cultured in vitro presented irregular polygonal shape,being closely connected in clusters,and surrounded by a small quantity of fibroblasts.The majority of cells were successfully labeled with CK7 and Lgr5,with expression primarily observed in the cytoplasm.The positive rate of cells successfully labeled with CK7 and Lgr5 was over 90％,indicating a high purity of epithelial cells.Conclusion The vestibular epithelial cell culture system was established in vitro through tissue-block culture technology,thereby providing a suitable cell model for further research on the vestibular functions and pathophysiological mechanisms of inner ear diseases and hair cell regeneration.
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