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Optimized Generation of Primary Human Epithelial Cells from Larynx and Hypopharynx: A Site-Specific Epithelial Model for Reflux Research

Laryngopharyngeal reflux (LPR) induces a differential damage effect on several anatomic sites within the larynx and hypopharynx; therefore, an in vitro model is needed for each anatomic site. This study aimed to establish a primary culture method for human laryngeal and hypopharyngeal epithelial cel...

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Autores principales: Mo, Ting-Ting, Tan, Jia-Jie, Wang, Mei-Gui, Dai, Yuan-Feng, Liu, Xiong, Li, Xiang-Ping
Formato: Online Artículo Texto
Lenguaje:English
Publicado: SAGE Publications 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7103601/
https://www.ncbi.nlm.nih.gov/pubmed/30917697
http://dx.doi.org/10.1177/0963689719838478
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author Mo, Ting-Ting
Tan, Jia-Jie
Wang, Mei-Gui
Dai, Yuan-Feng
Liu, Xiong
Li, Xiang-Ping
author_facet Mo, Ting-Ting
Tan, Jia-Jie
Wang, Mei-Gui
Dai, Yuan-Feng
Liu, Xiong
Li, Xiang-Ping
author_sort Mo, Ting-Ting
collection PubMed
description Laryngopharyngeal reflux (LPR) induces a differential damage effect on several anatomic sites within the larynx and hypopharynx; therefore, an in vitro model is needed for each anatomic site. This study aimed to establish a primary culture method for human laryngeal and hypopharyngeal epithelial cells derived from multiple anatomic sites. Surgical mucosa specimens were treated with a two-step enzymatic strategy to establish a primary culture. Of the 46 samples, primary cultivation was achieved successfully with 36 samples, and the positive ratio was 78.3%. In addition, flow cytometry revealed that these primary cells were epithelial cells with a purity of 94.9%. The proliferative ability was confirmed by positive staining for Ki-67. Laryngeal and hypopharyngeal epithelial cells from multiple sites exhibited similar epithelial morphology and positive cytokeratin expression. These cells can be cultured to passage 4. In summary, we successfully established the in vitro epithelial model of larynx and hypopharynx subsites, which may potentially be used as a platform for reflux research, especially for site-specific damage effect.
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spelling pubmed-71036012020-04-03 Optimized Generation of Primary Human Epithelial Cells from Larynx and Hypopharynx: A Site-Specific Epithelial Model for Reflux Research Mo, Ting-Ting Tan, Jia-Jie Wang, Mei-Gui Dai, Yuan-Feng Liu, Xiong Li, Xiang-Ping Cell Transplant Original Articles Laryngopharyngeal reflux (LPR) induces a differential damage effect on several anatomic sites within the larynx and hypopharynx; therefore, an in vitro model is needed for each anatomic site. This study aimed to establish a primary culture method for human laryngeal and hypopharyngeal epithelial cells derived from multiple anatomic sites. Surgical mucosa specimens were treated with a two-step enzymatic strategy to establish a primary culture. Of the 46 samples, primary cultivation was achieved successfully with 36 samples, and the positive ratio was 78.3%. In addition, flow cytometry revealed that these primary cells were epithelial cells with a purity of 94.9%. The proliferative ability was confirmed by positive staining for Ki-67. Laryngeal and hypopharyngeal epithelial cells from multiple sites exhibited similar epithelial morphology and positive cytokeratin expression. These cells can be cultured to passage 4. In summary, we successfully established the in vitro epithelial model of larynx and hypopharynx subsites, which may potentially be used as a platform for reflux research, especially for site-specific damage effect. SAGE Publications 2019-03-27 2019-05 /pmc/articles/PMC7103601/ /pubmed/30917697 http://dx.doi.org/10.1177/0963689719838478 Text en © The Author(s) 2019 http://creativecommons.org/licenses/by-nc/4.0/ This article is distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 License (http://www.creativecommons.org/licenses/by-nc/4.0/) which permits non-commercial use, reproduction and distribution of the work without further permission provided the original work is attributed as specified on the SAGE and Open Access pages (https://us.sagepub.com/en-us/nam/open-access-at-sage).
spellingShingle Original Articles
Mo, Ting-Ting
Tan, Jia-Jie
Wang, Mei-Gui
Dai, Yuan-Feng
Liu, Xiong
Li, Xiang-Ping
Optimized Generation of Primary Human Epithelial Cells from Larynx and Hypopharynx: A Site-Specific Epithelial Model for Reflux Research
title Optimized Generation of Primary Human Epithelial Cells from Larynx and Hypopharynx: A Site-Specific Epithelial Model for Reflux Research
title_full Optimized Generation of Primary Human Epithelial Cells from Larynx and Hypopharynx: A Site-Specific Epithelial Model for Reflux Research
title_fullStr Optimized Generation of Primary Human Epithelial Cells from Larynx and Hypopharynx: A Site-Specific Epithelial Model for Reflux Research
title_full_unstemmed Optimized Generation of Primary Human Epithelial Cells from Larynx and Hypopharynx: A Site-Specific Epithelial Model for Reflux Research
title_short Optimized Generation of Primary Human Epithelial Cells from Larynx and Hypopharynx: A Site-Specific Epithelial Model for Reflux Research
title_sort optimized generation of primary human epithelial cells from larynx and hypopharynx: a site-specific epithelial model for reflux research
topic Original Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7103601/
https://www.ncbi.nlm.nih.gov/pubmed/30917697
http://dx.doi.org/10.1177/0963689719838478
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