EPC2-hTERT cells were used for ALI experiments (25 (link)). Cells were grown submerged in KSFM media on a 0.4 μm filter (Corning Life Sciences) for three days to reach confluency (protocol schematic, Figure 2A ). Cultures were then switched to high-calcium concentration KSFM media (1.8 mM Ca++) for 5 days. Media was removed from the upper chamber to promote epithelial differentiation for days 7 to 10 (25 (link)).
Protocol detail
Differentiation of EPC2-hTERT Airway Epithelial Cells
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Corresponding Organization : University of Pennsylvania
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Variable analysis
independent variables
- Culture media (KSFM vs high-calcium concentration KSFM)
- Epithelial differentiation
- Cell line (EPC2-hTERT cells)
- Cell culture substrate (0.4 μm filter)
- Cell culture duration (3 days to reach confluency, 5 days in high-calcium media, 3 days in ALI condition)
- Positive control: Not mentioned
- Negative control: Not mentioned
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