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Lung Toxicity Service

Assays

  • LDH release
  • ATP assay
  • MTT assay
  • TEER

Models

  • Human small airway epithelial cells on air liquid interface (ALI) (24 well)

Timeline

  • 2-4 months

Newcells has developed an air-liquid interface (ALI) culture as a valuable tool for modelling the small airways of the lung in vitro and evaluating the lung toxicity of potential drug candidates

Lung toxicity and lung fibrosis occur as a result of inhalation of particles, pathogens and therapeutics but also following repeated administration of systemic and oral drugs.

Any epithelial damage or loss of epithelial barrier integrity leads to an increased susceptibility to infection, an impairment of gas exchange and triggers an immune response leading to inflammation, tissue damage and possibly lung toxicity.  In vitro epithelial cell culture models such as our SAEC model provides predictive data  enabling accurate predictions of the potential cellular toxicity of therapeutics by assessing multiple epithelial damage parameters.

Service outputs

  • LDH release assay
  • ATP activity quantification
  • MTT activity
  • TEER

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Small Airway Epithelial Cells immunostained to detect junctional protein ZO1, a marker for tight junctions

Multiple readouts

All major cell types

of the lung epithelia

24-well format

Air-liquid interface

Lung Toxicity Assay

Accelerate your lead compound selection by evaluating their safety and lung toxicity profile in a relevant model of the lung small airways

1.

Accurately investigate response to epithelial damage

2.

Easily evaluate lung safety/toxicity in vitro

3.

De-risk drug discovery

Service overview Close Open

Cultured on 3D permeable supports, our fully differentiated, polarised, human small airway epithelial cell (SAEC) model allows the addition of xenobiotics to either the apical or basolateral compartments to evaluate how they affect the epithelial barrier integrity  and if they trigger cytokine release in response to cell damage.

As a standard, we measure four readouts our assays to determine drug-induced lung toxicity. As an example, Puromycin, an antibiotic with known toxic side effects in the lung,  triggers  a dose dependent response as measured by TEER, LDH release and ATP/MTT activity.

 

Puromycin induces a dose-dependent response as measured by A) TEER, B) LDH release, C) cellular ATP activity and D) formazan formation. After stimulating Newcells’ SAEC-ALI model with specified concentrations of Puromycin for 72 hours, resultant cell and epithelium damage show a dose-dependent decrease in TEER, which is associated with an increase of LDH release. Data shows a dose-dependent reduction in ATP activity and cell viability.
Assay design
Models

Primary small airway epithelial cell model with an ALI (air liquid interface).

The model includes all major cell types of the lung epithelium:

  • Basal cells
  • Club cells
  • Goblet cells
  • Ciliated cells
Assay format

24-well

Species

Human

Assay readout
  • TEER
  • Cell viability (LDH, ATP, MTT)
Biological variation

3 human healthy donors (recommended)

Technical replicates

3 technical replicates per condition (recommended)

Assay treatment window

72 hours

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