I am trying to culture MDA MB 231 cells in culture flasks. I need to know if the cells are contaminated by yeast. The cells are base adherent. The color of media is mostly transparent with a shade of pale yellow (not really different than uncontaminated culture flask). Media composition is Hyclone DMEM with 10% FBS and 1% penstrap. The cells have multiplied over a period of 24 hours despite small dots in the attached pictures. The suspect (small white dots) are visible clearly in both magnified images although better in 40X. the small dots arent moving at all. Maybe 1 out of 10,000 are propogating in small space. I suspect that the small dots maybe yeast but I am not sure since I have never seen them in past.
Hello, in your images it looks like the brightvspots are inside the cells. They look like lipid droplets. The cells may be stressed by culture conditions. I suggest checking the pH of the incubated medium. The culture medium recommended by the ATCC is L-15 in air (no CO2). Have you tried Leibovitz-15 medium? https://www.atcc.org/products/all/HTB-26.aspx#culturemethod
what you have are dead cells mostly floating on top of the adherent cells. your DMEM media with co2 should be still OK. Leibovitz-15 medium is expensive and make it hard if you are comparing results with other cells like MCF-7.
use fresh media and make sure that your serum is heat inactivated. if your media was produced 6 months ago, I suggest to add fresh sodium pyruvate and glutamax. let me know how it goes with you. also check that your incubator temperature and co2 level are constant
Do NOT try Leibovitz-15 if you culture your cells in CO2 incubator. Use DMEM instead.
Thanks for all your inputs. The culture conditions of the cells have been approved by a professor of mine at Stanford who lent me these cells. I have not tried Leibovitz-15 medium yet. The previous time I tried to revive a new vial of same cell line, they grew just fine using same media, FBS and penstrap under same culture conditions.
I just checked the newly revived culture today that used a completely new bottle of medium and FBS. The vial was revived 3 days ago and none of the cells are attached or grown. I have attached the pictures along with this comment.
1) Does it look like there might be a mycoplasmic contamination?
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