Xi Ren 1 , 2 , Philipp T Moser 1 , 2 , Sarah E Gilpin 1 , 2 , Tatsuya Okamoto 1 , 2 , Tong Wu 1 , 2 , Luis F Tapias 1 , 2 , 3 , Francois E Mercier 1 , 2 , Linjie Xiong 1 , 2 , Raja Ghawi 1 , 2 , 4 , David T Scadden 1 , 2 , 5 , Douglas J Mathisen 2 , 3 , Harald C Ott 1 , 2 , 3 , 5
Bioengineered lungs produced from patient-derived cells may one day provide an alternative to donor lungs for transplantation therapy. Here we report the regeneration of functional pulmonary vasculature by repopulating the vascular compartment of decellularized rat and human lung scaffolds with human cells, including endothelial and perivascular cells derived from induced pluripotent stem cells. We describe improved methods for delivering cells into the lung scaffold and for maturing newly formed endothelium through co-seeding of endothelial and perivascular cells and a two-phase culture protocol. Using these methods we achieved ∼75% endothelial coverage in the rat lung scaffold relative to that of native lung. The regenerated endothelium showed reduced vascular resistance and improved barrier function over the course of in vitro culture and remained patent for 3 days after orthotopic transplantation in rats. Finally, we scaled our approach to the human lung lobe and achieved efficient cell delivery, maintenance of cell viability and establishment of perfusable vascular lumens.