Dissociation, Cellular Isolation and Initial Molecular Characterization of Neonatal and Pediatric Human Lung Tissues.

Human lung morphogenesis begins by embryonic life and continues after birth into early childhood to form a complex organ with numerous morphologically and functionally distinct cell types. Pulmonary organogenesis involves dynamic changes in cell proliferation, differentiation, and migration of specialized cells derived from diverse embryonic lineages. Studying the molecular and cellular processes underlying formation of the fully functional lung requires isolating distinct pulmonary cell populations during development. We report now novel methods to simultaneously isolate four major pulmonary cell populations from pediatric human lung. Cells were dissociated by protease digestion of neonatal and pediatric lung and isolated on the basis of unique cell membrane protein expression patterns. Epithelial, endothelial, non- endothelial mesenchymal and immune cells were enriched by fluorescence activated cell sorting. Dead cells and erythrocytes were excluded by 7-AAD uptake and glycophorin-A (CD235a) expression ...