We culture our cells in DMEM High glucose with 10% FBS and ABAM. However, previously we were using High Glucose w/o L-Glutamine and now the media contains L-Glutamine. Even after reviving our older frozen stocks, the cells appear stressed, right after transfection. They appear just fine before transfection though. Could the change in media composition be the issue? Please help!!
transfection is probably not a gentle experience for the cells ! How do you know your cells are stressed?
it would probably be better to use the same media for transfection and culture, however HEK are quite robust. Is the DNA you are transfecting clean ? what protocole did you use to purify it (no phenol!)? Did you try other type of transfection (lipofection)?
Transfection usually stress the cells to some extent. You might see your HEK cells getting more round. They should look fine after 24hr-48hr post-transfection. Use a reporter gene (such as fluorescent protein) to know if your normal looking cells are effectivley transfected.
I usually culture my cells in DMEM but transfect in OptiMEM.
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