Figure 3
Analysis of respiratory epithelial cell-like differentiation in expanded CB-CD34+ cells. A-C: Immunofluorescence analysis of surfactant protein-C expression in expanded CB-CD34+ cells, day 21, showing SP-C immunoreactivity (red) in cells cultured in DEX medium (A) and basic medium (B). C: Isotype IgG control. DAPI counterstain (blue). Scale bar = 20 μm. D. RT-PCR analysis of surfactant protein-C gene expression in expanded CB-CD34+ cells exposed to Dex at concentrations ranging from 10-7M to 10-5M. Shown are the results of two representative isolates. Highest SP-C expression of SP-C (327 bp) is seen in the presence of Dex at 10-5M. Positive (human lung) and negative controls (omission reverse transcriptase, H2O control) were included. The housekeeping gene GAPDH (407 bp) was included as loading control. E: RT-PCR analysis of surfactant protein-C gene expression in expanded CB-CD34+ cells. Shown are the results of one representative isolate. Expression of SP-C is seen in the presence of dexamethasone (DEX, 10-5M) after 15, 18 and 21 days of culture. Controls were as described under D.