What are the consequenses of too low/high seeding densities of Caco-2 cells, or other intestinal epithelial cell lines, in transwell culture systems? Or does cell concentration at seeding not affect the resulting monolayer's barrier function? Is it just a matter of time before the barrier integrity is established?
There are debates on how confluent should be the cells on transwell filter. However , the number of seeded cell on insert has been declined over time in published papers i.e from 300000 till 50000 in 12 well plates e.g. The high density couses multilayer formation and very low some how effect the polarization and final establishment of integrity. It is not only time but other factors e.g number of cells and medium and FBS % play roles.
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