Pneumologie 2015; 69 - A45
DOI: 10.1055/s-0035-1556637

hiPSC derived endothelial cell types from scalable cultures for biofunctionalization and tissue engineering

R Olmer 1, 2, M Szepes 1, S Menke 1, 2, M Pflaum 1, 2, S Schmeckebier 1, 2, I Gruh 1, U Martin 1, 2
  • 1Leibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), Hannover Medical School, REBIRTH – Center for Regenerative Medicine
  • 2Biomedical Research in Endstage and Obstructive Lung Disease Hannover (BREATH) Member of the German Center for Lung Research (DZL)

Introduction: Applications like full endothelialisation of gas exchange membranes in extracorporal membrane oxygenation (ECMO) devices for improved hematocompatibility, cell therapy of pulmonary hypertrophy or tissue engineering require large numbers of (patient-specific) endothelial cells (ECs). The isolation of ECs from peripheral blood or explanted vessels is well established however especially cells from older individuals show a limited proliferation capacity. Patient specific ECs from pluripotent stem cells (hiPSCs) might be an alternative suitable cell source. The opportunity to generate large amounts of undifferentiated hiPSC in defined media under scalable conditions allows for the generation of cell numbers in dimensions which are suitable for envisioned applications. By differentiation of these well monitored cell populations a virtually unlimited number of (autologous) ECs may become available for disease modelling, tissue engineering approaches and biofunctionalization of ECMO devices.

Methods & Results: Application of VEGFA and a small molecule Gsk3ß inhibitor under defined medium conditions resulted in up to 46% of CD31pos cells from hPSC cultures. Via FACS the amount of CD31pos cells could be increased to > 90%. Sorted hPSC derived CD31pos cells could be further expanded, formed networks after seeding to matrigel cultures and showed homogenous Dil-Ac-LDL uptake. Utilization of these cell populations for seeding of membranes for extracorporal oxygenation resulted in a homogenous cell layer on the hollow fiber membrane 48h after seeding. Addition of hPSC derived endothelial cells to tissue engineered cardiac constructs resulted in complex network structures distributed throughout the construct.

Conclusion: Resulting patient- (and lung disease-) specific iPSC-derived ECs will represent a novel cell source for disease modelling or biofunctionalization of gas exchange membranes as well as for vascularization of tissue engineered constructs. In addition, TALEN-based gene correction in iPSCs might enable novel concepts of ex vivo gene therapy for respiratory diseases.

*Presenting author