The Hela cell was a gift from other lab, and it grows well in the other lab. However, after several passages, the cell look like the following pic. The media I am using is DMEM+L- glutamine+10%FBS+ antibiotics. By the way, the media of cell is pretty clear. I don’t know the cause.
Hello Lei,
which passage are these cells? Normally primary cells like HeLa you can only passage sometimes, not often. For HeLa some people say about 20-25 passages, for example here: http://rudra.hubpages.com/hub/hela-cell-culture
In my opinion most primary cells can only passage up to 5 times, like it is written here for example: http://encodeproject.org/ENCODE/protocols/dataStandards/Cell_Culture_Guidelines.pdf
So I think your cells die.
If the photo is anything to go on, it looks like your cells have a bacterial contamination. Hela cells should not have that level of background debris in the media
Hi Sven,
thanks for your suggestions. actually, it is Hela cell line, not primary cell. And it grows quite well in the other lab. We prepare our DMEM medium from Gibico powder, and there is something floating in the medium after filtered probably due to the glass bottle dried in the oven. I dont know the reason, but after we got the cell, the background debris appear after the first passage, and cell appeareance starts to change after the next 3~4 passage.
we want to try it with comercial Hyclone medium, and i hope the cell cuture problem will be solved.
Maybe you can give cells to the other lab also? Or get a vial from the other lab? When they then also not grown in your lab you have to check why.
Hi,
What I can notice from your pic is that you have some kind of precipitate, and my guess is that you have a contamination in your culture (looking at the bottle of medium does not always reflect the actual state). There are two more things: I always add sodium pyruvate and and adjust sodium bicarbonate to the one optimal for HeLa cells.
Hope that this helps!
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