Gene induction during differentiation of human pulmonary type II cells in vitro

Studies of human fetal lung in explant culture and in isolated epithelial cells have demonstrated that both glucocorticoids and cyclic AMP promote differentiated alveolar type II cell phenotype as assessed by ultrastructural morphology and surfactant production. This project profiles changes in gene expression associated with hormone induced differentiation. Undifferentiated human fetal lung (13-20 wk) epithelial cells were cultured in serum-free medium (control) or with dexamethasone/8-Bromo cyclic AMP/isobutylmethylxanthine (DCI) to promote type II cell differentiation. RNA from five sets of experiments (10 samples) was evaluated using the U133A Affymetrix GeneChip set. Keywords: Hormone treatment Cells were isolated from 13 individual lungs and the cells from each were cultured in the absence (control) and presence of DCI dexamethasone (10 nM)/8-Br-cyclic AMP (0.1 mM)/isobutylxanthine for 72 h. RNA was isolated from the control and treated cells of each prep. For the 5 sets of microarray experiments (10 total chips, 2 for each control and treated pair), RNA from 2 of the cell preps was analysed as individual microarray comparisons. RNA from 11 cell preps was pooled for the other 3 experiments (4, 4, and 3 cell preps) to provide control and DCI-treated RNA pools.