Hello Every body, i have a problem in cell culture of Caco-2, after each passage the cell count were reduced till in passage 6 there was no cell in our flask and we saw a white layer in our flask , would You please tell me is it normal or our cells had contamination? I also put the photo of each passage. and if someone work with this cell would you please please send me the precedure of working.
thanks a lot
Hi Salmeh, I am sorry to hear that your Caco-2 cells had some problem. I have worked with several kinds of epithelial cells, including Caco-2. It is possible that your cells have some contamination (or fungi or yeast or else). Since this cell line is not difficult to culture. Normally I sub-culture the cell cells every 2-3 days, and I used this medium: Caco-2 cells were cultured in DMEM (Gibco) supplemented with 10% (v/v) FBS, 1 mM glutamine, 1% (v/v) non-essential amino acids (Gibco), penicillin (50 units/mL) and streptomycin (50 µg/mL).
Generally, in tissue culture if your flask has a white layer on the top of your cells and cell strat to detached from the container that means that the flask is contaminated. just check the turbidity of the medium and test it by microbiological culture.
Also, I would recommend checking the medium and the supplementations. Always use antibiotics if multiple contaminations occurred.
@Guangzhi Zhang @ Saeed Mohammad Kabrah thanks alot for your comments
Have you noticed there are CaCO-2 debris in your mycoplasma culture? :P
I'm sorry to say that but the common source of mycoplasma is cross-contamination from previously infected cells/media/sera. I would strongly recommend you to discard all your contaminated flasks, solutions and vials. Use always sterile-filtered solutions
Dear Sylvia
thank alot for Your Comment, we know they were not normal because we think that cell source from where we got the cells had been contaminated with mycobacterium and as you said our reference picture is what is in ATCC site.
it seems that non of your picture resembles attached caco-2. I also culture cac0-2 in my lab. Attached herewith the photomicrograph of my cac0-2 for your reference. Hope it may help..
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