HI,
I am working on breast cancer and I\m trying to use MCF12F as normal breast cell line. I purchased it from ATCC and Iam having problem growing it. It dosent seem to attach and the few cells that have attached are floating after few days. The cells are like this and have added everything to the medium as recommended by ATCC. They replaced the cell twice for me but still its the same. Does anyone have this problem. Suggestions are welcome.
The non-tumorigenic epithelial cell line MCF12F are cultured in DMEM/F12 (1:1, GIBCO). Medium was supplemented with 10% fetal bovine serum, 10 μg/ml insulin, 0.5 μg/ml hydrocortisone, 20 ng/ml recombinant EGF and antibiotics. ATCC recommended the addition of cholera toxin, however we do not used this supplement in the medium growth
Yes I added the above said components to it. I dont know if I should add cholera toxin or not.
I havent coated the plate with collagen or matrigel but can try. The media composition is DMEM/F12 (1:1, GIBCO). Medium was supplemented with 10% fetal bovine serum, 10 μg/ml insulin, 0.5 μg/ml hydrocortisone, 20 ng/ml recombinant EGF and antibiotics. ATCC recommended the addition of cholera toxin, however we do not used this supplement in the medium growth
Dear Jaya,
You have done according to the procedures recommended. try to check whether there is no contamination?
good luck
Iam sure there is no contamination. Medium is clear and not stinking. The problem is that the cells are not attaching when it platted the primary vial from ATCC for what so ever reason which I dont know.
Dear Jaya,
I came across your thread, as I have the same problems with MCF-12 cells. Did you ever find a solution?
I culture in the same conditions (but with Horse serum instead of bovine, and I do add cholera toxin)
Many thanks,
Stacey
MCF-12F was isolated as a floating culture; that's what the "F" stands for in MCF-12F. When I used to grow them, they never adhered, nor did I think they were supposed to adhere. But ATCC site says they are an adherent cell line, lol.... but, I was able to propagate and treat them just fine as a floating culture.
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