Technique development for airway epithelial biology
We use primary cultured cells from human and mouse (as well as others) nose or lung to study the multiciliated airway epithelium, and we are always looking for ways to enhance our ability to manipulate and characterize these cells. Cultures are initiated by isolating stem cells from airway tissues (obtained from human donors by nasal or bronchial brushing or during transplant or other surgery) and seeding them on porous Transwell membranes. Cells proliferate to confluence under submerged conditions, then differentiate at an air-liquid interface (ALI), which is created by feeding the cells only from below the Transwells. We have developed and optimized ALI culture methods to permit the assessment and manipulation of epithelialization, ciliogenesis and PCP using confocal, electron and timelapse microscopy, FACS and lentiviral gene transfer.