title: Combinatorial enzymatic digestion with thermolysin and collagenase type I improved the isolation and culture effects of hair cell progenitors from rat cochleae.
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.
authors.affiliation: Department of Otolaryngology-Head and Neck Surgery, Xijing Hospital, Fourth Military Medical University, Xi'an, 710032, China.
authors.forename: Yong-Li
authors.initials: YL
authors.lastname: Song
authors.affiliation: Department of Otolaryngology-Head and Neck Surgery, Xijing Hospital, Fourth Military Medical University, Xi'an, 710032, China.
authors.forename: Ke-Yong
authors.initials: KY
authors.lastname: Tian
authors.affiliation: Department of Otolaryngology-Head and Neck Surgery, Xijing Hospital, Fourth Military Medical University, Xi'an, 710032, China.
authors.forename: Wen-Juan
authors.initials: WJ
authors.lastname: Mi
authors.affiliation: Department of Otolaryngology-Head and Neck Surgery, Second Affiliated Hospital, Xian Jiaotong Medical University, Xi'an, 710004, China.
authors.forename: Peng
authors.initials: P
authors.lastname: Han
authors.affiliation: Department of Otolaryngology-Head and Neck Surgery, Xijing Hospital, Fourth Military Medical University, Xi'an, 710032, China.
authors.forename: Zhong-Jia
authors.initials: ZJ
authors.lastname: Ding
authors.affiliation: Department of Otolaryngology-Head and Neck Surgery, Xijing Hospital, Fourth Military Medical University, Xi'an, 710032, China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Qiu
authors.affiliation: Department of Otolaryngology-Head and Neck Surgery, Xijing Hospital, Fourth Military Medical University, Xi'an, 710032, China.
authors.forename: Fu-Quan
authors.initials: FQ
authors.lastname: Chen
authors.affiliation: Department of Otolaryngology-Head and Neck Surgery, Xijing Hospital, Fourth Military Medical University, Xi'an, 710032, China. qiujh@fmmu.edu.cn.
authors.forename: Jian-Jua
authors.initials: JJ
authors.lastname: Qiu
mesh_terms: D000818:Animals; D001489:Basilar Membrane / Q000166:cytology; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002469:Cell Separation / Q000379:methods*; D003051:Cochlea / Q000166:cytology*; D017364:Collagenases / Q000494:pharmacology*; D004847:Epithelial Cells / Q000166:cytology; D006198:Hair Cells, Auditory / Q000166:cytology* / Q000187:drug effects; D017207:Rats, Sprague-Dawley; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects; D013820:Thermolysin / Q000494:pharmacology*
chemical_list: D017364:Collagenases; C417249:collagenase 1; D013820:Thermolysin
keywords: Cell culture; Enzymatic digestion; Epithelial cells; Hair cell progenitors