Hi all, I observed black spots in my cultured macrophages. The complete culture medium I used includes DMEM, FBS, penicillin-streptomycin, and amphotericin B. However, the cells still got contaminated. I filtered the medium using a 0.22 µm CA membrane filter. I also disinfected the incubator, fumigated the lab, and replaced the incubator filters, but these black spots still won’t go away. Does anyone know which step went wrong or what else I could do? Thank you so much in advance!
In this situation, it is possible that there is an issue with the serum. Before use, did you notice any abnormalities in the serum, such as flocculent precipitates?
Generally, the components of serum precipitates can be simply classified as follows, and they typically do not affect cell culture. However, given your current situation, it is suspected that the serum may be problematic. We recommend centrifuging the serum at 400g, collecting the supernatant, and then using it to prepare the culture medium.
After preparing the complete medium, filter it and incubate without cells to observe whether the contamination persists. If contamination is still present, you may consider performing mycoplasma detection. If contamination is confirmed, it may be necessary to purchase new serum.
This answer is provided by MedChemExpress (MCE) Technical Support. [email protected]
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