I'm culturing mouse mucosal and gingival epithelial cells and noticed these cell aggregates. Does anybody know what they may be? Is this the early stages of a fungal contamination? I thawed and cultured these epithelial cells in epi media and noticed these odd looking cells after two days of growth. I went ahead and split the cells and took these pictures after another two days of culturing in an incubator at 37 degrees Celsius at 5% CO2. These aggregates (or possibly fungi) increased in numbers, but I don't see any filamentation or hyphae. Any thoughts?
I don't think so, although hard to be certain. Typically fungal contaminations have visible processes (mycelia) which these appear to lack, as you've noted. To me this looks more like clumped cell over-growth due to the high confluence of the monolayer. Unless your protocol indicates otherwise I would suggest a more harsh split ratio/lower seeding density to prevent cell reaching confluence, although experience suggests mouse fibros like to grow that way.
If these blebs are in fact free-floating, or the media has turned more pink than usual (assuming standard Phenol Red-containing media), I would consider reviving another vial or testing for fungi.
https://handling-solutions.eppendorf.com/cell-handling/contamination/scientific-background/fungal-contamination/
If your tissue isdissociated to single cell suspensions before culture, another approach maybe helpful: selective removal of the cell foci. These foci (clumps) of cells may represent subpopulations of cells. You can try cold detachment/dissociation enzyme or EDTA treatment while observing the dish or flask. When the clumps retract, gently rinse them off and discard (or dissociate and culture separately to see what you get).
- Badges
- Science topic
- Similar topics
- Phytochemistry
- Extracts