I am a newcomer to cell culture, and it is the first time to culture a HEK293 cell line that stably expresses Kv4.3, but I feel that the cell state is very poor, the cell membrane is incomplete after recovery(Picture1), and a large number of dead cells and cell debris accumulate and attach to the cell surface during the subsequent culture(Picture2), and the cells grow slowly.
Cell Culture Method
Medium：DEME High Clucose + 10% FBS + 1% Penicillin-Streptomycin (100×)
3.5 cm Petri dish；Medium change every day
I would like to ask if anyone has raised this kind of cell, or what is wrong with cell culture in the photo, thank you very much!!!!
I don't have any experience with the channel protein that you are working with, but I can tell you that HEK 293 cells are not very adherent. They tend to detach easily. Maybe ease up on the media changes and let them lay down completely. These cells shouldn't need daily media changes unless the channel expression requires it. I split 293 cells twice a week at most and leave them alone in between passages.
Are these cells freshly thawed, or have they been in culture for some time? Are they under selection? How did you introduce the channel gene (lentivirus, plasmid transfection)?
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