Hi,
I am starting to work with immortalized MEFs, and can't find any information about the density of MEFs at 100% confluency.
Can anyone help me with that? In the end the question is: how many cells are there in a 10cm dish at 100%?
I culture them in DMEM with 10% FBS and 1% Pen/Str/Gln.
Thanks!
The cell number in the plate would depend on the size of cells. Further, the size would depend on culture conditions. Assuming that everything is "normal", the estimate would be between 2-5 million cells/plate.
MEFs can vary quite a bit depending on how they were immortilized and what other mutations they may have picked up along the way. If MEFs are contact dependent still, you'll likely be on the low end of Divaker's range. Are your cells growing in a single layer or 3-dimensionally?
Mark, not sure how they grow, to be honest, I don't have much experience with cell culture. It looks like they generally grow in a single layer, but tend to partially sit on top of each other.
MEFs are immortalized or transformed? If cells in culture are exhibiting a transformed phenotype (serum independence and the lack of contact inhibition) due to a spontaneous transformation, the number could be much higher. Could you include a photograph of cells under your culture conditions?
Dear Divaker, as far as I know, they are immortalized with SV40. I will be able to make a photograph tomorrow, but can't promise high quality :(
I agree with Divaker. If your MEFs still exhibit contact inhibition, then 2-5 million per 10cm dish is a reasonable estimate. If they stratify, all bets are off on how many can pack into the culture vessel.
I'm guessing you have the answer by now, but i get between 2 and 3 million per plate.
I use fresh MEFs up to P5 and also home made 3T3 MEFs (after P20) from different KO mice. Just at conflency, the number of cells in a 60mm dish (56cm2) is near 3.106. But you can let grow cells and go up 4 more cells during one week after confluency. The problem is this induce scenecence of cells and cells seems fine but only 30% are able to grow after passaging.
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