- I want to do expansion for my A549 cells to bigger plate after selection with (Ab) drug there is a lot of drug resistant cells colonies but I lost MOST of them when I tried to harvest them by trypsinization(that take longer time than usual and still cells attached 10 min!) and centrifugation...what could be the reason behind this...!!
Are you sure that the adherence of the cells wasn't affected by the transfection/drug selection? I haven't had A549 attach to cell culture flask walls before, but sometimes the transfection reagent used can lead to high levels of adherence (we use this one, and it works: https://altogen.com/product/a549-transfection-reagent-lung-carcinoma-ccl-185/). Otherwise, I would look at the effects of the transfected cargo (if you introduced plasmids) and see in previous research if it resulted in some strange effects.
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