I have a student who has tried on numerous occasions to propagate MB-231s without success. So far we purchased three different batches of these cells from ATCC… We sequentially cultured the cells from each vial in the recommended L-15 medium + 10% FBS, and incubated them without exposure to CO2. On three occasions, however, there was formation of a “milky” or a cloudy layer in the medium after 24 hours of incubation … Ultimately the cells, fail to adhere on the bottom of the flasks and, die.
Hi Lehlohonolo- we have always cultured our MB-231s in RPMI 1640 + 10% FBS + pen/strep at 5% CO2. This line attaches and grows very well. Have you contacted ATCC about possible contamination on their end? Hope this helps, and good luck!
If you have contamination, I think of bacteria because you said cloudy medium, it is possible that an aliquote of medium, serum or other reagents were contaminated and you used the same solution in all the three batches.
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