Abstract
The high incidence of hearing loss in human combined with the lack of hair cell regeneration in mammalian cochleae had got the attention to manipulate stem/progenitor cells to participate in hair cell regeneration for years. Cochlear progenitor cells are considered as the best candidate for hair cell regeneration. However, there is not any effective and feasible way to separate hair cell progenitors from rat cochleae, yet. In this study, we tried to isolate single epithelial cells from rat basilar membrane by combinatorial enzymatic digestion with thermolysin and collagenase type I. The results showed that the harvested single cells gave rise to otospheres with features of stem cells and could be induced to differentiate into hair cells. Significantly, more otospheres of epithelial origin were obtained by digesting with thermolysin and collagenase type I. The combinatorial enzymatic digestion would be a potential method for hair cell progenitor isolation and culture with broad applications.
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This work was supported by the National Natural Science Foundation of China (grant no. 81271069) and the National Natural Science Foundation of Major Projects Overseas of China (grant no. 81120108008)
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All procedures concerning the animals in this study were approved by the Institutional Animal Care and Use Committee of the Fourth Military Medical University and were in compliance with the National Institute of Health Guide for the Care and Use of Laboratory Animals.
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Editor: Tetsuji Okamoto
Yong-li Song and Ke-yong Tian contributed equally to this work.
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Song, Yl., Tian, Ky., Mi, Wj. et al. Combinatorial enzymatic digestion with thermolysin and collagenase type I improved the isolation and culture effects of hair cell progenitors from rat cochleae. In Vitro Cell.Dev.Biol.-Animal 52, 537–544 (2016). https://doi.org/10.1007/s11626-015-9998-4
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DOI: https://doi.org/10.1007/s11626-015-9998-4