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in this video we will discuss how the tier values are correlated with air liquid interface culture morphology ali cultures generated using the pneumocult culture system will display an optimal morphology in early passages roughly p2 to p5 good to acceptable performance is achievable in passages p5 to p8 respectively beyond p8 the differentiation potential declines and the ali differentiation assay is not recommended at week 1 post ali the cells will become tightly packed and bound by tight junctions creating a high barrier function subsequently tier values will peak at week 2 post ali the pseudostratified epithelium will begin to form and cells determined for differentiation will begin to stand upright into their columnar morphologies tier values will fall into an ideal tier range as paracellular and trans cellular movements begin to take place at week 3 post ali as the cellular differentiation into ciliated cells and goblet cells continues to develop the tier values remain wit