Induction of mouse lung injury by endotracheal injection of bleomycin

Fiorenza Orlando, Chiara Paolini, Silvia Agarbati, Cecilia Tonnini, Antonella Grieco, Chiara Capelli, Martino Introna, Mauro Provinciali, Armando Gabrielli, Gianluca Moroncini

Research output: Contribution to journalArticlepeer-review


Pulmonary fibrosis is a hallmark of several human lung diseases with a different etiology. Since current therapies are rather limited, mouse models continue to be an essential tool for developing new antifibrotic strategies. Here we provide an effective method to investigate in vivo antifibrotic activity of human mesenchymal stromal cells obtained from whole umbilical cord (hUC-MSC) in attenuating bleomycin-induced lung injury. C57BL/6 mice receive a single endotracheal injection of bleomycin (1.5 U/kg body weight) followed by a double infusion of hUC-MSC (2.5 x 105) into the tail vein, 24 h and 7 days after the bleomycin administration. Upon sacrifice at days 8, 14, or 21, inflammatory and fibrotic changes, collagen content, and hUC-MSC presence in explanted lung tissue are analyzed. The injection of bleomycin into the mouse trachea allows the direct targeting of the lungs, leading to extensive pulmonary inflammation and fibrosis. The systemic administration of a double dosof hUC-MSC results in the early blunting of the bleomycin-induced lung injury. Intravenously infused hUC-MSC are transiently engrafted into the mouse lungs, where they exert their anti-inflammatory and antifibrotic activity. In conclusion, this protocol has been successfully applied for the preclinical testing of hUC-MSC in an experimental mouse model of human pulmonary fibrosis. However, this technique can be easily extended both to study the effect of different endotracheally administered substances on the pathophysiology of the lungs and to validate new anti-inflammatory and antifibrotic systemic therapies.

Original languageEnglish
Article numbere58922
JournalJournal of Visualized Experiments
Issue number146
Publication statusPublished - Apr 2019


  • Bleomycin
  • C57BL/6 mice
  • Endotracheal injection
  • Human umbilical cord mesenchymal stromal cells (hUC-MSC)
  • Immunology and Infection
  • Issue 146
  • Lung injury
  • Tail vein infusion

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)


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