Do Airway Epithelium Air–Liquid Cultures Represent the In Vivo Airway Epithelium Transcriptome?

Abstract

Human airway epithelial cells cultured in vitro at the air-liquid interface (ALI) form a pseudostratified epithelium that forms tight junctions and cilia, and produces mucin. These cells are widely used in models of differentiation, injury, and repair. To assess how closely the transcriptome of ALI epithelium matches that of in vivo airway epithelial cells, we used microarrays to compare the transcriptome of human large airway epithelial cells cultured at the ALI with the transcriptome of large airway epithelium obtained via bronchoscopy and brushing. Gene expression profiling showed that global gene expression correlated well between ALI cells and brushed cells, but with some differences. Gene expression patterns mirrored differences in proportions of cell types (ALIs have higher percentages of basal cells, whereas brushed cells have higher percentages of ciliated cells), that is, ALI cells expressed higher levels of basal cell-related genes, and brushed cells expressed higher levels of cilia-related genes. Pathway analysis showed that ALI cells had increased expression of cell cycle and proliferation genes, whereas brushed cells had increased expression of cytoskeletal organization and humoral immune response genes. Overall, ALI cells provide a good representation of the in vivo airway epithelial transcriptome, but for some biologic questions, the differences between in vitro and in vivo environments need to be considered.

Collective migration and cell jamming in asthma, cancer and development

“…To answer this question, Park et al assessed both the dynamic and structural signatures of the jamming transition in human bronchial epithelial cells (HBECs) that were derived from non-asthmatic and asthmatic donors (Park et al, 2015b). To recapitulate in vitro the differentiation and repair processes that occur in the maturing or injured airway epithelium in vivo, they used the well-established air-liquid interface (ALI) culture approach to recapitulate well-differentiated pseudostratified HBEC layers as in vivo airway epithelium (Dvorak et al, 2011;Whitcutt et al, 1988).…”

Non-animal models of epithelial barriers (skin, intestine and lung) in research, industrial applications and regulatory toxicology

“…Placing PBECs at an ALI in the presence of retinoic acid causes the epithelial cells to form a pseudostratified structure with basal cells supporting ciliated cells and mucus-producing goblet cells (Pezzulo et al, 2010), as occurs in vivo. Transcriptomic studies have shown that ALI cultures provide a good representation of the in vivo airway epithelial transcriptome (Dvorak et al, 2011;Pierrou et al, 2007). Many in-house airway epithelial models based on primary human cells have been developed and used (Bérubé et al, 2010;Gray et al, 1996;Xiao et al, 2011).…”

Molecular Mechanisms of Metaplasia, Differentiation and Hyperplasia of Goblet Cellin Allergic Asthma

“…However, in vitro cultures do not always reflect physiologic conditions in vivo [104,105]. The pathological changes in asthma affect the epithelium morphology and function.…”

“…Several oxidative stress studies revealed that endogenous antioxidant enzymes were decreased in the peripheral tissues of asthmatic adult patients relative to the control subjects, indicating that mitochondrial dysfunction and oxidative stress is present in asthmatic adults [115]. However the relation of those changes and mucus production in vitro and in vivo events [105] istotally unknown.Thus, additional work is needed to fully answer this questionsfor better therapy in the control of mucus hypersecretion. Production of TSLP and cumulative effects in the development of asthma, leading to goblet cell responses and epithelium damage.…”