Hi. I am trying to culture 16hbe14o- cells (human bronchial epithelial cells) on 24/6 well transwell inserts. The cells grow well in T75 flasks but I am struggling to get a mono layer on the inserts. What seeding density should be used? For how long should the cells be cultured? and how often should the media be changed? We are not using any coatings.
Shyam Nyati
How long do you wait after plating before you stain the cells? If you are doing migration assay 8 hours or so might be sufficient, for invasion assays you might increase the time (to 24 hours) and the number of cells as well. I am guessing that you plate the cells (in the chamber) in serum free media while a chemo attractant is media with serum or your growth factor?
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