Intratracheal administration of solutions in mice; development and validation of an optimized method with improved efficacy, reproducibility and accuracy

Charlotte E Pelgrim*, Ingrid van Ark, Thea Leusink-Muis, Maike A D Brans, Saskia Braber, Johan Garssen, Ardy van Helvoort, Aletta D Kraneveld, Gert Folkerts

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

3 Citations (Web of Science)


Animal models are still vital in the field of respiratory disease research. To improve the accuracy and consistency of the dose of specific compounds administered specifically in the respiratory tract, it is important to optimize and to compare the technique to currently available techniques. In this study, an optimized intubation-mediated intratracheal administration (IMIT) technique is described and compared to oropharyngeal aspiration (OA). Adult female Balb/c mice were treated with Evans Blue using IMIT or OA and sacrificed after a short recovery to observe the distribution of solutions throughout the lungs. Additionally, mice were treated with increasing doses of lipopolysaccharide (LPS) or saline to compare efficacy of both techniques. Inflammatory cell numbers in bronchoalveolar lavage were quantified 24 h post-administration. Evans Blue staining revealed a more homogeneous distribution and less variability among animals treated using IMIT as compared to OA. Higher inflammatory cell numbers were observed in IMIT mice compared to OA mice after exposure to vehicle or the lowest LPS concentration. This study shows that the optimized IMIT is superior to OA with regards to efficacy, reproducibility and accuracy. This IMIT method can be deployed to refine 3R animal welfare aspects of the experimental design and improve the reproducibility of respiratory disease mouse models.

Original languageEnglish
Article number107156
Number of pages7
JournalJournal of Pharmacological and Toxicological Methods
Publication statusPublished - 4 Feb 2022


  • Animals
  • Bronchoalveolar Lavage Fluid
  • Disease Models, Animal
  • Female
  • Intubation, Intratracheal
  • Lipopolysaccharides
  • Lung
  • Mice
  • Mice, Inbred BALB C
  • Reproducibility of Results
  • Research Design
  • Oropharyngeal
  • LUNG
  • Intratracheal
  • Instillation
  • Netherlands
  • Intubation
  • Methods

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