I have worked with HEK293 FT cells for many years. Recently, they have mysteriously started to look abnormal (see attached). After being passaged and grown in complete DMEM media (10% FBS, Pen/Strep, L-Glut, Geneticin) for ~3 days, instead of being fully confluent, they become rather elongated, skinny and sparse with the media colour remained mostly red. When they were left to grow for another day without changing media, they started to die off. However, if I passaged them, they would grow normally for 2 days. Despite this issue, when cells were seeded and transfected, they expressed okay. Myoplasm was tested negative. I have tried cleaning the incubator, made up fresh media and recovered a new vial of cells but the issue remains unresolved. Could unstable temperature or low CO2 percentage in the incubator cause this kind of cell growth?
It could be a problem with your CO2, if it isn't injecting or reading properly. Low CO2 makes it so that your culture media can't maintain the proper pH and will cause cells to sicken and die. It could also explain why they are ok for a little while after being passaged into fresh media.
I'm not sure what else the problem could be. They look a bit like cells do when you infect them with a lentivirus, but that doesn't seem likely based on your explanation.
Thanks Theresa. The cells continue to grow badly in the incubator after 3 days. The CO2 and temperature logs were normal over the past weekend, unless the readings are off. The thermometer I put in is showing 38deg rather than 37deg but that wouldn't make cells unhappy, would it? I have now grown more cells in other incubators as a test.
What is the passage number of your cells, those exhibited altered morphology?
Do you have the facilities to check the expression of EMT markers; if yes, you may perform those analyses
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