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Isolation, culture, and identification of ceruminous gland cells

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Abstract

External auditory canal (EAC) stenosis or atresia usually requires a skin graft to repair, but due to the lack of a graft containing functional glands, postoperative complications such as infection and eczema are common. The aim of this study was to isolate and characterize seed cells for the construction of tissue engineered EAC skin containing ceruminous gland by isolating and cultivating cells of ceruminous gland. In this study, EAC skin samples were harvested from adult goats for ceruminous gland cell isolation. Cell morphology and proliferation rates, expression of CK7, CK8, CK18, and CK19 (glandular cell specific-markers), and secretion of β-defensin-1, lysozyme, and polysaccharides were evaluated at different passages to verify the presence of ceruminous gland cells and determine whether function and proliferation potential were maintained. Ceruminous glands were successfully isolated and extracted from goat EAC skin. Furthermore, the isolated glandular cells maintained robust proliferation potential, exhibited high expression of CK7, CK8, CK18, and CK19, and vigorously secreted β-defensin-1, lysozyme, and polysaccharides in this culture system. However, expression of glandular cell specific-markers and secretory function gradually declined with increasing passage number, indicating dedifferentiation of the subcultured ceruminous gland cells after five passages. In conclusion, ceruminous glands were successfully isolated, cultured, and expanded from goat EAC skin using the serumcontaining culture system. Importantly, the isolated glandular cells retained robust proliferation potential and maintained their phenotype and function in early passages (P1–P3), indicating the method’s potential application for ceruminous gland regeneration.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (82172105, 31700837, 81771014).

Funding

This work was supported by the National Natural Science Foundation of China (31700837, 81800920, 81771014).

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Author notes

  1. Jun Wang and Aijuan He have contributed equally to this work.

Authors and Affiliations

  1. Department of Otorhinolaryngology, Head and Neck Surgery, Division of Life Sciences and Medicine, The First Affiliated Hospital of USTC,, University of Science and Technology of China, Hefei, 230001, Anhui, People’s Republic of China

    Jun Wang

  2. Department of Facial Plastic and Reconstructive Surgery, Eye & ENT Hospital of Fudan University, Shanghai, People’s Republic of China

    Aijuan He, Yaying Zhu & Tianyu Zhang

  3. Hearing Medicine Key Laboratory, National Health and Family Planning Commission, Shanghai, People’s Republic of China

    Tianyu Zhang

  4. Department of Plastic and Reconstructive Surgery, Shanghai 9th People’s Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai Key Laboratory of Tissue Engineering, Shanghai, People’s Republic of China

    Guangdong Zhou

  5. ENT Institute, Eye & ENT Hospital of Fudan University, 83 Fenyang Road, Shanghai, 200031, People’s Republic of China

    Jun Wang & Tianyu Zhang

Authors
  1. Jun Wang
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  2. Aijuan He
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  3. Yaying Zhu
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Correspondence to Tianyu Zhang.

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The authors have no conflict of interest to declare.

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This study was approved by the Ethics Committee and the Institutional Review Board of the Eye & ENT Hospital of Fudan University (Shanghai, P.R. China). All animal experiments were carried out in accordance with the U.K. Animals (Scientific Procedures) Act, 1986 and associated guidelines, EU Directive 2010/63/EU for animal experiments.

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Wang, J., He, A., Zhu, Y. et al. Isolation, culture, and identification of ceruminous gland cells. J Mol Histol 53, 227–237 (2022). https://doi.org/10.1007/s10735-021-10040-y

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  • DOI: https://doi.org/10.1007/s10735-021-10040-y

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