In the past few weeks I've run into some familiar and unfamiliar problems with my mouse small intestine organoids grown in IntestiCult media.
I've attached photos and I'm wondering if anyone has seen anything like them before.
The first two photos are heavily contaminated with what might be yeast?
The next two have debris that initially looks like single cells from the organoids in both the matrigel and the surrounding media. However, if given time, they will multiply and make the media quite turbid, which makes me suspect them as a foreign contaminant. Additionally, at higher magnification I cannot morphologicaly distinguish these from very small organoid bodies.
The final two are a mystery to me. I've seen my organoids "shed" before where they slough off some cells from the outside before falling apart and dying. These however appear to be intact and relatively healthy organoids with large clusters of cells clinging to the outside. What I can't tell is whether those cell clusters are organoids or a foreign cell type contaminating the culture.
Any insight in these scenarios would be much appreciated!
It appears that yes first 2 are contaminated with yeast. Not sure about the next 2: possibly artifacts in Matrigel? The 5th could be bacterial contamination stuck to organelles after a media change?
I dont think it is coming from matrigel. This looks like yeast if it is upto 2X magnification or coccus bacteria if it under higher magnification.
You need to have better sterilization.
Hi Kimberly,
looks yeast to me. Maybe using Fungizone for the next cultures might help to avoid it. Keeping the instruments clean also will help (pipettes mainly).
good luck!