Gene delivery to the lung in vitro – reducing the number of transgenic animals

Dr. Marius Hittinger, Prof. Dr. Claus-Michael Lehr & Dr. Henrik Groß,
PharmBioTec GmbH,
Saarbrücken, Germany


New therapeutic approaches for the treatment of severe lung diseases (e.g. cystic fibrosis) are increasingly being tested on genetically modified laboratory animals. The relevance of such experiments for the human body is often questionable.

The project "Gene therapy of the lung in vitro - reducing the use of transgenic animals to a minimum" used human systems to investigate how gene therapy of the human lung can be simulated in vitro to reduce the use of animal experiments in this rapidly growing field of research.

To this end, various mechanisms such as particle deposition, uptake into the various lung cells and efficacy were mimicked. In addition to cell lines, primary alveolar cells from the human lung were also used for this purpose. Primary autologous coculture of the lower airways was of great importance for the investigation of new pulmonary formulations.

In the experimental setup, the upper airways were mimicked by the Calu-3 cell line, while the lower airways were simulated with primary alveolar lung cells and cocultures. The influence of macrophages in the lower airways or mucus from the upper airways on the efficacy of new forms of therapy was investigated.



Fig. 1:
Different in vitro models of the alveolar and bronchial region of the lung will be investigated at the air-liquid interface and at submerged culture conditions. Effects on transfection efficacy as a consequence of the model will be determined.

The knowledge gained is immediately transferred between Saarland University, the Helmholtz Institute for Pharmaceutical Research and PharmBioTec GmbH. Research in the field of gene delivery is carried out at all locations so that new findings can be directly incorporated into similar work.


M. Hittinger et al., Cell and tissue-based in vitro models for improving the development of oral inhalation drug products, Eur. J. Pharm. Biopharm. (2017), dx.doi.org/10.1016/j.ejpb.2017.02.019

Vukosavljevic B, Hittinger M, Hachmeister H, Pilger C, Murgia X, Gepp MM, Gentile L, Huwer H, Schneider-Daum N, Huser T, Lehr CM, Windbergs M. Vibrational spectroscopic imaging and live cell video microscopy for studying differentiation of primary human alveolar epithelial cells. J Biophotonics. 2019 Jun;12(6):e201800052. doi: 10.1002/jbio.201800052. Epub 2019 Feb 20. PMID: 30597770.

Sapich S, Hittinger M, Hendrix-Jastrzebski R, et al. Murine Alveolar Epithelial Cells and Their Lentivirus-mediated Immortalisation. Alternatives to Laboratory Animals. 2018;46(2):73-89. doi:10.1177/026119291804600207


PharmBioTec GmbH, Saarbrücken, Germany


01/2016 - 04/2017