Establishment and long-term culture of human cystic fibrosis endothelial cells
Endothelial cell (EC) dysfunction has been reported in cystic fibrosis (CF) patients. Thus, the availability of CF EC is
paramount to uncover mechanisms of endothelial dysfunction in CF. Using collagenase digestion, we isolated cells from
small fragments of pulmonary artery dissected from non-CF lobes or explanted CF lungs. These cells were a
heterogeneous mixture, containing variable percentages of EC. To obtain virtually pure pulmonary artery endothelial cells
(PAEC), we developed an easy, inexpensive, and reliable method, based on the differential adhesion time of pulmonary
artery cells collected after collagenase digestion. With this method, we obtained up to 95% pure non-CF and CF-PAEC.
Moreover, we also succeed at immortalizing both PAEC and CF-PAEC, which remained viable and with unchanged
phenotype and proliferation rate over the 30th passage. These cells recapitulated cystic fibrosis transmembrane
conductance regulator expression and functions of the parental cells. Thus, we isolated for the first time endothelial cells
from CF patients, providing a valuable tool to define the emerging role of EC in CF lung and vascular disease.