Characterizing Exogenous Cell Engraftment for Cystic Fibrosis Cell Therapy

Abstract

Cell therapy is a potential method for treating cystic fibrosis (CF), although cell engraftment into the highly resistive, pseudostratified human bronchial epithelium (HBE) is a major challenge. Transient epithelial injury is an approach to improve cell uptake, but the extent of injury required and the ability of cells grown by alternative methods to engraft are poorly understood. Our overarching hypothesis is that compounds such as polidocanol, an FDA approved drug used to sclerose varicose veins, can increase cell engraftment by targeting luminal barrier cells. To test this hypothesis, we engrafted exogenous, adenovirus‐EGFP transduced primary human airway epithelial cells into well‐differentiated air‐liquid interface (ALI) HBE cell cultures following treatment with varying doses of polidocanol. Quantitation of cell engraftment demonstrated that 0.125% polidocanol increased engraftment without causing overall ALI cell culture lysis. Further confocal microscopy characterization of the polidocanol injury indicates that engraftment does not require epithelial denudation to the level of the basal lamina. Primary HBE ALI cultures were then injured with polidocanol and the recovery of transepithelial electrical resistance was tracked over time after addition of no cells, conventionally‐grown cells, or cells grown with the conditionally reprogrammed cell (CRC) method. Cell engraftment expedites epithelial restoration, and CRC‐grown cells encourage recovery sooner than conventionally‐grown cells. CF ALI HBE cell cultures engrafted with non‐CF cells were tested for functional CFTR recovery in Ussing chambers. CRC HBE cells engraft, persist and recapitulate CFTR function more effectively than conventionally‐grown cells. Our results suggest that transient polidocanol injury and the CRC culture method may be a viable approach to enhance cell engraftment for CF cell therapy.Support or Funding InformationSupported by CFF grants RANDELXX015 and BOUCHE15R0CF2015, and NIH grants U01HL111018 and P30DK065988.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.