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Small Airway Epithelial Cell Model

Model formats

  • Small Airway Epithelial Cell Model
  • 24-well air-liquid interface (ALI) culture

Cell types

  • Basal cells
  • Club cells
  • Goblet cells
  • Ciliated cells

Species

  • Human (Healthy donor)

A robust, physiologically relevant air-liquid interface (ALI) lung model of the small airways to enable scientific research and drug discovery

Newcells Biotech’s human small airway epithelial cell (SAEC) air-liquid interface model closely recapitulates the epithelial physiology of the lung. Derived from differentiated small airway basal cells, our SAEC model comprises the main epithelial cell types namely basal, club (Clara), goblet and ciliated cells. With an established epithelial barrier, active mucus production and functional cilia, our air-liquid inferface (ALI) SAEC model is an invaluable tool for drug discovery.

The small airways of the lung are the terminal bronchioles, vital for the conduction of air from the larger airways to the alveolar region for gas exchange. Unlike the larger airways, the small airways are non-cartilaginous, with a diameter < 2 mm, making them a key site of airway resistance. The presence of excessive mucus within the small airways can often lead to airway obstruction such as that observed in chronic obstructive pulmonary disease (COPD), asthma, and other respiratory diseases. Recent studies have also suggested a role of the small airways in the development and progression of lung fibrosis. As the study of the small airways in vivo is limited due to their fragility, size and location, our in vitro human lung small airway epithelial cell model provides a powerful tool for the study of lung physiology, drug toxicity and response to epithelial damage.

We currently offer a range of analytical readouts in line with project requirements, example readouts include;

  • TransEpithelial Electrical Resistance (TEER)
  • Cytokine release
  • Gene expression modifications
  • Immunocytochemistry
  • Cell viability & compound toxicity
Small Airway Epithelial Cells immunostained to detect for Basal cell (KRT5, green), Club cell (CC10/SCGB1A1, pink) and Junctional protein (ZO1, red) markers.

Human primary

low passage cells

Three human donors

Mimics in vivo tissue

air-liquid interface

Human lung small airway epithelial cell (SAEC) model

A robust, physiologically relevant air-liquid interface (ALI) lung model of the small airways to enable research and drug discovery

1.

Physiologically relevant air-liquid interface (ALI) small airway model

2.

Contains all key lung epithelial cell types

3.

Testing of compounds toxicity on the small airways

Applications for small airway epithelial cell model Close Open

Air-liquid interface (ALI) culture holds enormous potential for respiratory research, with a wide range of applications including:

  • Modelling of respiratory diseases
  • Toxicity testing of therapeutic compounds

Our team have over 10 years of experience working with lung epithelial cells across a wide range of assays and would be happy to discuss your project needs

Human Small Airway Epithelial Cell Model Close Open

Formation of a 3D Pseudostratified Epithelium

Immortalised cell lines and primary cells derived from animals models are commonly used, however, the data they generate lack relevance to human physiology. In order to recapitulate the pseudostratified mucociliary phenotype observed in vivo, human lung small airway epithelial cells need be cultured at the air-liquid interface.

For our ALI-SAEC model, primary human small airway basal cells are expanded in culture flasks before being seeded onto permeable Thincert supports containing culture medium in both the apical and basal compartments. The basal cells are then expanded until confluent prior to being subjected to “air-lift”, i.e. with medium only supplied to the basal chamber to create the air-liquid interface. This configuration mimics the conditions found in human and drives differentiation towards a mucociliary phenotype, forming a 3D pseudostratified epithelium containing club cells in greater abundance than goblet cells. Full differentiation takes 21-28 days and cultures are used from 21-days onwards.

Expression of Junctional Genes & Proteins

This model is further characterised by the development of a functional epithelial barrier, as indicated by the expression of tight junction proteins and the development of high transepithelial electrical resistance (TEER) as shown by staining of the junctional proteins ZO1 and E-Cadherin. ZO1 and E-Cadherin protein expression, in conjunction with TEER data, confirms cells are polarised and successfully differentiated. Gene expression levels of junctional genes (TJP1, JAM-A, CDH1) can also be tracked throughout the differentiation period.

SAEC cell culture was immunostained for the tight junction marker ZO1 and the adherens junction marker ECadherin

Cell-Specific Gene Expression

As the basal cells differentiate, we quantify the expression of cell-specific genes as an indirect assessment of basal cell differentiation by qPCR. This also confirm the presence of all relevant cell types following 21 days in culture.

Cell-Specific Protein Expression

MUC5B and MUC5AC are mucin proteins expressed by goblet cells in both the upper and small airway, however their ratio differs depending on the location. MUC5B is predominantly expressed in the small airways. Validation data collated from the SAEC model suggests a higher expression of MUC5B than MUC5AC,  similar to the expression pattern observed in vivo.

Newcells’ small airway epithelial cell model expresses key cell specific proteins. (A) ZO1 staining shows presence of epithelial tight junctions. Presence of club cells (B), goblet cells (C) and ciliated epithelial cells (D) as indicated by CC10, MUC5B and acetylated tubulin (AcT) expression.

 

Immunostaining of SAEC model for cell-specific proteins

Validation of Increase in Ciliary Beating Frequency during SAEC differentiation

Newcells’ SAEC model cultured in ALI conditions has successfully demonstrated increase in cilia expression during differentiation. Data from the CiliaBeat software analysis confirms an increase in ciliary beat frequency  and area of the apical cell surface covered by active cilia from day 14 post-airlift onwards aligning with physiological characteristics.

(A) Heatmaps produced by CiliaBeat software showing percentage area covered by active cilia and their ciliary beat frequency (B) and (C) Increase in ciliary beat frequency and percentage area covered by active cilia at different timepoints post air-lift.

Cilia beating

Brightfield recording of fully differentiated Small Airway Epithelial Cells in air-liquid interface (ALI) cell culture with functional beating ciliary movements.

Images

Small Airway Epithelial Cells immunostained to detect Acetylated Tubulin (AcT), a marker for Ciliated cells
Small Airway Epithelial Cells immunostained to detect junctional protein E-Cadherin, a marker for Adherens Junctions
Small Airway Epithelial Cells immunostained to detect junctional protein ZO1, a marker for Tight Junctions
Small Airway Epithelial Cells immunostained to detect junctional protein ZO1, a marker for Tight Junctions
Small Airway Epithelial Cells immunostained to detect junctional protein ZO1, a marker for Tight Junctions
Small Airway Epithelial Cells immunostained to detect mucin proteins MUC5AC (green) and MUC5B (red), which are markers for Goblet cells.
Small Airway Epithelial Cells immunostained to detect for Basal cell (KRT5, green), Club cell (CC10/SCGB1A1, pink) and Junctional protein (ZO1, red) markers.
Brightfield Image of Small Airway Epithelial Cells in air-liquid interface (ALI) cell culture.
Small Airway Epithelial Cells immunostained to detect for Basal cell (KRT5, green), Club cell (CC10/SCGB1A1, pink) and Junctional protein (ZO1, red) markers.

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