Table 3 HTF isolation using 10% DMEM
Description of each isolation step | Passage number | Culture dish used | Time needed to reach monolayer | Features of obtained monolayer |
|---|---|---|---|---|
Biopsy cutting and setting up of the isolation | 0 | Well of 12-well plate | 13.3 ± 3.1 days | Co-culture of mostly epithelial cells and single HTFs |
Cell detachment using trypsin and cell scraper | 1 | One 25 cm2 T-flask | 13 days- failure | Few clusters of epithelial cells and single HTFs |
Medium change from DMEM to FGF-enriched EMEM | - | One 25 cm2 T-flask | 15.5 ± 2.1 days | Mostly HTFs and single epithelial cells |
Cell detachment using trypsin | 2 | Two 25 cm2 T-flasks | 2 days | 99–100% of vimentin-positive HTFs |
Trypsinization and cryopreservation of HTFs | - | - | - | - |