Hi everyone!
So I am having a hard time with my HEK293 cell line. I have been successful in culturing them in the past, but for some reason for the last couple of months I've been having problems keeping them alive. I use DMEM with 10% FBS, PBS as a wash and trypsin to unadhere the cells. They are kept in an incubator at 37 degrees and 5 % CO2. After passaging the cells, they seemed to be adhered and happy. I checked about 2 days later and they were happily adhered. 2 days after that, I checked on my cells and they were completely detached and floating in the media. This has happened to a few of my lab mates as well and we can't figure out what the issue is. Is there advice or tips/tricks to passaging these cells successfully?
Thank you!
You could try e.g. to change cell culture plates for those ones that enhance cell adhesion. The following discussion on RG can help: https://www.researchgate.net/post/Cells_detaching_in_culture
Hello Rivas,
I think your cells already get confluence over 80% that may be the reason for detachment of cells. One another reason like starvation of cells also responsible for the same. I am agreed with @Yvonne Baumer's comments.
Try to replace media on 2nd day and try to check growth and subculture at 70-80% confluence.
Good luck.
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