I am having trouble with the MCF10A cells that I am working on. Some are sticking to the bottom of the plates but they are not growing and there is always debris in the plate after 3-4 times of PBS washing them. Is there a way to make these cells grow in plates I would love to do something with them?
Hello Ryan Rahim
What is the growth medium you are using?
For MCF-10A cells the medium preparation is as follows:
DMEM/Hams F12, supplemented with 2mM L-glutamine, 0.5mM sodium pyruvate, 20ng/ml human Epidermal Growth Factor (hEGF), 100ng/ml cholera toxin, 10µg/ml bovine insulin, 500ng/ml hydrocortisone, 5% horse serum and 1% Penstrep.
At what passage number are your cells growing ? These cells grow fast at the early passages and then they slow down with late passages in culture.
You also need to check for mycoplasma contamination.
Best.
Here is the medium for MCF-10A in our lab. The components will be added into 10% FBS DMEM medium. Please see the attachment.
Malcolm Nobre I am using this media recipe but I feel like the cells are still not attaching to the plate and I am using a plate that has membrane lining on the bottom of the plate for better attachments. I think whoever froze the cells was not stored properly but I have been through over 5 thawings. Do you have any recommendations?
Doan Huu Nhat Binh How come the media recipe provided does not contain choleratoxin?
If you think that the cells were not stored properly, you can thaw a vial from another source ( from another laboratory) to prove your point.
Did you check the viability percentage of the cells on thawing?
Malcolm Nobre I will keep you updated, I will try that. Thank you so much.
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