I had got a vial of KB cell lines from abroad, preserved in dry ice. The packaging and transport were in proper form. I revived the cells in a T-75 flask and they started to attach after 10 days in big clumps. I changed the medium and then split the cells into a T-25 flask. However, they refused to attach and died within 2-3 days. Can anyone explain why this happened?
Hello.
I have never worked with this cell line but I can suggest something about your culture conditions. It is very strange that these cells require 10 days for attach. So, please control the composition of medium (in particular % FBS) and cell density (number of cells for cm2), Second, when they attach, before to perform the split, wait until the cells reach the best confluency.
The follow link speak about composition of medium and cell culture conditions of your cell line
https://www.phe-culturecollections.org.uk/products/celllines/generalcell/detail.jsp?refId=94050408&collection=ecacc_gc
Good luck
Thank you Sir for your suggestion. I had used 20% FBS for revival of cell lines and the cell density was a billion cells in a T-75 flask.
I think you have to try with 10% FBS and 1,5x106 cells in T-75 flask.
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