Fig 1.
Phase contrast images of A549 monolayers.
Images show the differences in morphology of 25 day continuous culture in DMEM (A) or Ham’s F12 (B).
Fig 2.
Cell counts, viability and cell diameters from A549 cell cultures at day 25. A: cell yield per unit area based on cell counting using Trypan blue dye exclusion (n = 3) and B: cell numbers as measured by automated cell counting (n = 5). C: Cell viability based on DAPI dye exclusion (n = 5). D: cell diameter using automated image analysis (n = 5). Data are plotted as mean ± SD; statistical analyses in A, B and D used unpaired Student’s T Test. In C statistical testing showed no significant difference (NS).
Table 1.
Pathways Associated with Cell Cycle Control.
Fig 3.
RNA Microarray Heatmap of Gene Expression.
Hierarchically clustered (Euclidian similarity measure, clustered using Ward’s linkage rule) heat map of gene expression changes in A549 cells cultured in Ham’s F12 for up to 25 days; the heat map shows normalised intensity values of significant genes regulated up or down two-fold or more.
Table 2.
Pathways Associated with Apoptosis.
Table 3.
Pathways Associated with Senescence and Autophagy.
Table 4.
Pathways Associated with Epithelial and Endodermal Differentiation.
Table 5.
Pathways Associated with Lipid Synthesis and Metabolism.
Fig 4.
Shared up regulated (≥ 2 fold) gene expression of differentiated A549 with freshly isolated human primary ATII cultures.
Gene expression in A549 cells that were cultured for 7, 11, 18 or 25 days or primary ATII cells were compared to log phase A549 cells. The figure shows the number of shared up- (hatched bars) or down- (clear bars) regulated genes between the A549 cells and ATII cells over the A549 time course.
Table 6.
Pathways Considered to be Associated with ATII Phenotype.
Table 7.
Genes Related to ATII Differentiation.
Fig 5.
Oil-Red-O Staining of A549 Monolayers.
Phase contrast images of Oil-Red-O staining of lipid droplets in log phase A549 monolayers grown in Ham’s F12 medium in log phase (A), and cells in the same medium for 7 (B), Day 11 (C) and Day 18 (D) days.
Fig 6.
Electron Micrographs Showing Multilamellar Body Expression in A549 Cells.
Transmission electron micrographs of sections of A549 cells after 11 (A and C) or 21 (B and B) days of culture in Ham’s F12. Lipid bodies (Lb), mitochondria (m), nuclei (n) and multilamellar bodies (MLB) are identified. Scale bars = 500 and 1000 nm.