Pneumologie 2015; 69 - A47
DOI: 10.1055/s-0035-1556639

Differentiation of human embryonic and induced pluripotent stem cells into CFTR expressing cholangiocytes for drug screening and disease modeling

R Haller 1, 2, 3, L Engels 1, 2, 3, S Merkert 1, 2, 3, M Schubert 1, 2, O Ruth 1, 2, 3, M Ulrich 1, 2, 3
  • 1Leibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), Department for Cardiothoracic, Transplantation and Vascular Surgery, Hannover Medical School, Germany
  • 2REBIRTH Cluster of Excellence, Hannover, Germany
  • 3Biomedical Research in Endstage and Obstructive Lung Disease Hannover (BREATH), Member of the German Center for Lung Research (DZL), Hannover, Germany

Introduction:

Cystic fibrosis (CF) is the most common lethal monogenic recessive disease in the caucasian population. While the majority of patients die of respiratory failure, hepatobiliary disease is the third leading cause of death in CF. Over 1900 mutations are known so far in cystic fibrosis transmembrane conductance regulator (CFTR) gene, but the most common is the ΔF508 mutation, resulting in enhanced degradation of the CFTR protein. Currently approved CFTR modulators are only available for a minority of CF patients. In addition, these modulators possess varying potentials. Therefore finding new modulators is of exceptional importance and might be achieved by the establishment of new cell culture based platforms for drug screening and disease modeling.

Materials and Methods:

Human embryonic stem cell (hESC) and induced pluripotent stem cell (iPSC) are utilized for differentiation approaches. Cells are monitored for definitive endoderm and hepatic endoderm generation by flow cytometric analysis. Maturation of hepatoblasts into cholangiocytes is achieved by the addition of growth factors. CFTR expression is monitored via qRT-PCR and can be directly observed by application of hESCs and hiPSCs carrying a CFTR Tomato reporter construct.

Results:

The generation of definitive endoderm with hESCs as well as hiPSCs is with up to 97% highly efficient. Endodermal cells were further differentiated into hepatoblast and early cholangiocytes. Subsequently cholangiocytes were enriched and further maturated to gain up to 15% tomato-positive cells expressing CFTR.

Conclusion:

The protocol is a first step for hPSC derived cholangiocytes, which might represent a suitable basis for disease modelling and drug screening approaches of liver associated diseases. A halide sensitive reporter system carrying a CFTR Tomato reporter construct has been established as a basis for those drug screening approaches. These results might be translated to other organs e.g. the lung and facilitate CF research in the respiratory system.

*Presenting author