Hi everyone
Would anybody be able to tell me if it is possible to culture cystic fibrosis bronchial epithelial (CFBE) cells at ALI on transwell inserts? I have found plenty of literature that describes the successful culture of CFBE polarised monolayers using liquid-liquid interface (LLI) but nothing to date for ALI.
I have attempted to grow CFBE monolayers using 3 days under LLI and then switching to ALI. There is a monolayer of cells visible but the TEER is very low, even after 14 days of culture. Has anyone experience with this? Specifically, the optimal culture time under LLI before switching to ALI would be very helpful.
Thanks in advance,
Alan
Thank you both, Michelle and Rodolphe, these links should be very helpful!
I know I about a year late but were you able to get your ALI working properly? Your TEER could be low due to low seeding numbers or not having a fully differentiated mono layer (usually occurs in 16±2-3 days). Do you coat your inserts with collagen?
We usually keep LLI for 5 days after seeding on inserts and then from there feed regularly until we see ciliated cells.
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