Protein profile of well-differentiated versus un-differentiated human bronchial/tracheal epithelial cells

Wen-Kuan Liu,Duo Xu,Yun Xu,Shu-Yan Qiu,Li Zhang,Hong-Kai Wu,Rong Zhou

doi:10.1016/j.heliyon.2020.e04243

Wen-Kuan Liu, Duo Xu + Show 5 more

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https://doi.org/10.1016/j.heliyon.2020.e04243

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Abstract

Un-differentiated (UD) and well-differentiated (WD) normal human primary bronchial/tracheal epithelial cells are important respiratory cell models. Mature, WD cells which can be derived by culturing UD cells at an air-liquid interface represent a good surrogate for in vivo human airway epithelium. The overall protein profile of WD cells is poorly understood; therefore, the current study evaluated the proteomic characteristics of WD and UD cells using label-free LC-MS/MS and LC-PRM/MS. A total of 3,579 proteins were identified in WD and UD cells. Of these, 198 proteins were identified as differentially expressed, with 121 proteins upregulated and 77 proteins downregulated in WD cells compared with UD cells. Differentially expressed proteins were mostly enriched in categories related to epithelial structure formation, cell cycle, and immunity. Fifteen KEGG pathways and protein interaction networks were enriched and identified. The current study provides a global protein profile of WD cells, and contributes to understanding the function of human airway epithelium.

Highlights

Human airway epithelium is a pseudostratified layer consisting of basal cells, secretory cells, and columnar ciliated cells
We investigated the proteomic profiles of WD cells and UD cells using label-free Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS)
A total of 33 WD cell inserts were cultured at air-liquid interface (ALI) for 21 days

Summary

Introduction

Human airway epithelium is a pseudostratified layer consisting of basal cells, secretory cells, and columnar ciliated cells. The epithelium provides a critical interface between the body and the external environment (Crystal et al, 2008) This epithelial layer is known to be necessary for host defense against inhaled particles and microbes. In air-liquid interface (ALI) culture, un-differentiated normal human primary bronchial/tracheal epithelial (UD) cells can form a pseudostratified cell layer much like they do in vivo (Derichs et al, 2011). This well-differentiated normal human primary bronchial/tracheal epithelial (WD) cell model better mimics the in vivo environment than submersion culture which inhibits ciliogenesis and mucociliary movement (Min et al, 2016; Neugebauer et al, 2003). Structural and proteomic differences between WD and UD cells remains to be characterized

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