I'm a novice in mammalian cell culture. I'm culturing some MCF-7 cells (don't care about cells, they are not growing well because there's something wrong with the medium). I can frequently see this kind of things under the microscope in one of the culture dishes. They seem to have the same shape, but do not move. I'm wondering if they are contamination and what kind of contamination they are.
I don't think that's contaminated. Every once in a while you might get some lint from the cap of the cell culture flask inside your flask. But your cells certainly look like they're unhappy. Try a trypan blue stain to check viability.
I think the cells are not looking healthy. try with MEm with 10% FBS and !X sodium pyruvate
Hi Cheng ,
How long have the cells been seeded?which passage and density?
They indeed doesn't look healthy . what I propose is:
1- Thaw another vial , the cells could grow well and sometimes not , even the cryopreseved cells you have are the same.
2- seed at an appropriate density , so that the cells could proliferate well.(2500cell/cm2)
3-Try to find the appropriate culture media . (Eagle's Minimum Essential Medium, with 0.01 mg/ml human recombinant insulin and 10% fetal bovine serum )
4-Finally If you want to be sure if there is a contamination you can maybe do mycoplasma test
Hope it helps!
Good luck.
You can 1) keep them in medium without antibiotics for a while, 2) prepare microscope slides (for 100X), stained with dapi. Its hard to say, too big for bacterias, looks wierd, like a scratch on the lense. Do they appear after medium change?
About the MCF7, I agree with Amit kumar about the low density of your culture.
About the contamination,I don't think it's mycoplasme, but as say before, you can do a Dapi staining on your cell. About bacterial/fugus/yeast contamination, it's always difficult to know if it's contamination or not. In our lab, we have implemented a control system when we think it's could be a contamination : Centrifuge in a tube your culture medium (more the culture old more better), discard the supernatant but keep around 200ul. Up/Down to resuspend your pellet and spread 100ul on agar gelose and 100ul on potatoes gelose. Keep the gelose at 37°C until around 1 week. If you don't see colonies, it wasn't contamination. If you see colonies I suggest you a product name Virkon to clean all the culture room...
Good luck!
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