Gene expression profile of human lung epithelial cells with different culture methods.

The goal of our study is to establish a method to propagate human pneumocytes efficiently in a large scale. While human pneumectomy-derived lung epithelial cells (LECs) showed efficient long-term growth when co-cultured with irradiated NIH-3T3, they do not express molecular markers of mature pneumocytes or airway cells in this condition. Removal of feeders causes differentiation toward airway club cell-like phenotype, and an induction treatment with combination of small molecules and growth factors is required to differentiate them toward pneumocytes. To elucidate the underlying mechanisms, we analyzed gene expression profile of LECs with or without induction treatment. We also included LECs under coculture and the original frozen lung tissue, as a reference. Microarray experiments were performed as single-color hybridizations. Quality control and quantification of total RNA amount was assessed using an Agilent 2100 bioanalyzer (Agilent Technologies) and a NanoDrop 1000 spectrophotometer (Kisker).