Human airway epithelial cell cultures maintained at an air-liquid interface (ALI) recapitulate mucociliarydifferentiation found in vivo, regulating ion transport, mucin secetion and ciliated cell beating to reproducemucociliary clearance mechanisms characterisitic of the native human mucosa. These cultures are anexcellent model to study the role of CFTR, ENaC, nucleotide/nucleoside regulatory systems and theirintegrated function in health and disease as proposed in projects 1-4 of the current PPG, respectively. ATissue Procurement and Cell Culture Core was established at the University of North Carolina in 1984,under the auspices of the CF Foundation, to provide standardized cell cultures to CF researchers. The Corehas supported PPG projects since 2000 and has increased its output and capabilities to meet growingresearch demands. The Core routinely makes available cells and media that are unavailable and/orprohibitively expensive if purchased from commercial suppliers. The purpose of the present application is toprovide continuity in the supply of airway specimens and large numbers of human airway epithelial cellcultures to PPG investigators, and to support the use of cultures in mechanistic studies. To accomplishthese goals, we propose the following specific functions. 1) Tissue Procurement- Obtain nasal and bronchialtissues from normal, disease control, and CF humans as sources of airway epithelial cells and for acute exvivo experiments, biochemical studies, in situ hybridization and immunohistochemistry. Characterize donorswith respect to diagnosis, demographics, and clinical features relevant to research uses. 2) Cell Isolationand Culture- Isolate primary and passaged primary epithelial cells for distribution to project investigators.Maintain stocks of frozen cells and prepare media and substrates to support preparation of well-differentiatedairway epithelial cultures as dictated by investigator needs. 3) Genetic Mainipulation of Cell Cultures-Provide support for adenoviral and retroviral expression of molecules under study by project investigators.Through these functions, and in conjunction with other Cores in this application, the Cell Culture Core willsupport research to better understand mechanisms underlying normal airway surface liquid homeostatsis,which is essential for healthy lungs.
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