Temporal dynamics of ovine airway epithelial cell differentiation at an air-liquid interface

O’Boyle, Nicky; Sutherland, Erin; Berry, Catherine C.; Davies, Robert L.

doi:10.1371/journal.pone.0181583

Cited by 15 publications

(33 citation statements)

References 86 publications

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“…In particular, the transition of the undifferentiated monolayer to the fully differentiated epithelium must be well characterised over time. While the differentiation of human 27 , 39 and ovine 55 AECs over time has been previously studied, to date there has been no such characterisation of the temporal differentiation of bovine AECs. We have previously established optimal culture conditions for the growth and differentiation of bovine bronchial epithelial cells (BBECs) in a serum-free defined defined medium 56 .…”

Section: Introductionmentioning

confidence: 99%

Temporal differentiation of bovine airway epithelial cells grown at an air-liquid interface

Cozens

Sutherland

Marchesi

et al. 2018

Sci Rep

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There is an urgent need to develop improved, physiologically-relevant in vitro models of airway epithelia with which to better understand the pathological processes associated with infection, allergies and toxicological insults of the respiratory tract of both humans and domesticated animals. In the present study, we have characterised the proliferation and differentiation of primary bovine bronchial epithelial cells (BBECs) grown at an air-liquid interface (ALI) at three-day intervals over a period of 42 days from the introduction of the ALI. The differentiated BBEC model was highly representative of the ex vivo epithelium from which the epithelial cells were derived; a columnar, pseudostratified epithelium that was highly reflective of native airway epithelium was formed which comprised ciliated, goblet and basal cells. The hallmark defences of the respiratory tract, namely barrier function and mucociliary clearance, were present, thus demonstrating that the model is an excellent mimic of bovine respiratory epithelium. The epithelium was fully differentiated by day 21 post-ALI and, crucially, remained healthy and stable for a further 21 days. Thus, the differentiated BBEC model has a three-week window which will allow wide-ranging and long-term experiments to be performed in the fields of infection, toxicology or general airway physiology.

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Section: Introductionmentioning

confidence: 99%

Temporal differentiation of bovine airway epithelial cells grown at an air-liquid interface

Cozens

Sutherland

Marchesi

et al. 2018

Sci Rep

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“…Previous studies have placed this window between day 24-33 for human AECMs [32] and day 24-42 for ovine AECMs [44]. We have carried out comprehensive analysis of the differentiation of our model over an extended time-period and determine the window in which the bovine AECM were fully differentiated as between day 21-42 post-ALI.…”

Section: Discussionmentioning

confidence: 99%

“…The TEER is an important method for assessing the formation of junctional complexes between cells [54]. In our model, once confluency was reached, the TEER of the culture rapidly increased (Fig 3C) [25,44,55], however TEER of other AECMs have been shown to stabilised at the peak [32]. This variation in TEER over the course of culturing was not reflected in the presence of tight-junctional protein ZO-1, which was maintained at a stable level throughout the time-course.…”

Section: The Formation Of Tight-junctions Between Cells Creates a Phymentioning

confidence: 92%

“…Similarly, the transition of the model from undifferentiated cells to a fully differentiated epithelium must be well defined in order to determine the ability of the model to repair following damage. Although this has been achieved for human [28,32] and ovine AECMs [44], bovine AECMs have yet to be welldefined. We aimed to fully characterise the differentiation of BBECs grown at an air-liquid interface.…”

Section: Introductionmentioning

confidence: 99%

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Temporal differentiation of bovine airway epithelial cells grown at an air-liquid interface

Cozens

Sutherland

Marchesi

et al. 2017

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17The respiratory epithelium is exposed to assault by toxins and pathogens through the process 18 of inhalation, which has numerous implications on both human and animal health. As such, 19 there is a need to develop and characterise an in vitro model of the airway epithelium to study 20 respiratory pathologies during infection or toxicology experiments. This has been achieved 21 by growing airway epithelial cells at an air-liquid interface (ALI). Characterisation of ALI 22 models are not well-defined for airway epithelial cells derived from non-human species. In 23 this study we have fully characterised a bovine airway epithelial cell models (AECM) grown 24 at an ALI in relation to ex vivo tissue. The morphology of the model was monitored at three 25 day intervals, to identify the time-period at which the culture was optimally differentiated. derived from donor animals, our bovine AECM was shown to be highly representative of the 35 in vivo bovine bronchial epithelium and can be utilised in the study of respiratory 36 pathologies. 37 peer-reviewed)

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“…However, ALI approaches also proved useful to support differentiation and maintenance of in vivo-like functionality of epithelial cells that in vivo do not necessarily have apical access to ambient air, such as epithelia originating from the intestinal (Klasvogt et al, 2017;Nossol et al, 2011) and female reproductive tract (FRT) mucosa (for references see Table 1), on which we focus in this brief review article.…”

Section: Introductionmentioning

confidence: 99%

Air‐liquid interface cell culture: From airway epithelium to the female reproductive tract

Chen

Schoen

2019

Reprod Domestic Animals

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The air‐liquid interface (ALI) approach is primarily used to mimic respiratory tract epithelia in vitro. It is also known to support excellent differentiation of 3D multilayered skin models. To establish an ALI culture, epithelial cells are seeded into compartmentalized culture systems on porous filter supports or gel substrata. After an initial propagation period, the culture medium is removed from the apical side of the epithelium, exposing the cells to the surrounding air. Therefore, nutritive supply to the cells is warranted only by the basolateral cell pole. Under these conditions, the epithelial cells differentiate and regain full baso‐apical polarity. Some types of epithelia even generate in vivo‐like apical fluid or mucus. Interestingly, the ALI culture approach has also been shown to support morphological and functional differentiation of epithelial cells that are not normally exposed to ambient air in vivo. This review aims at giving a brief overview on the characteristics of ALI cultures in general and ALI models of female reproductive tract epithelia in particular. We discuss the applicability of ALI models for the investigation of the early embryonic microenvironment and for its implications in assisted reproductive technologies.

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Temporal dynamics of ovine airway epithelial cell differentiation at an air-liquid interface

Cited by 15 publications

References 86 publications

Temporal differentiation of bovine airway epithelial cells grown at an air-liquid interface

Temporal differentiation of bovine airway epithelial cells grown at an air-liquid interface

Temporal differentiation of bovine airway epithelial cells grown at an air-liquid interface

Air‐liquid interface cell culture: From airway epithelium to the female reproductive tract

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