Can this ALI system also be used for murine airway epithelial cells? And if yes do you have to change the protocol (as compared to human cells)? How many cells do you need per transwell?


Yes, the PneumaCult ALI system is compatible with murine airway epithelial cells. The protocol was optimised using commercially available human airway epithelial cells, so you might need to optimize the seeding density when you use murine airway epithelial cells. Starting with a range between 1X10^5 and 5X10^5 cells/cm^2 will likely provide a positive result. The cells should reach confluence after 1-4 days post-seeding at optimized seeding density. Optimised seeding density should also result in ALI differentiation exhibiting robust cilia beating and mucus secretion. Epithelium integrity and health can also be measured by transepithelial electrical resistance (TEER), which should be within 150 – 500 Ω∙cm^2 for high-quality ALI cultures.The seeding density also depends on the passage number, with cells at later passages typically resulting in lower-quality cultures. Given the anatomy differences between the human airway and the murine airway, you should expect a smaller proportion of goblet cells in the differentiated ALI cultures when you use murine airway epithelial cells.

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