I recently thawed THP-1 cells frozen at passage 20 in 2016 in 10% RPMI. After thawing, the cells appeared to recover successfully, and the next day I changed the media to remove the remaining DMSO. The cells were cultured in T-25.
On the 7thday, the two T-25 flasks showed turbid media. The other cells in the CO2 incubator with the THP-1 cells did not exhibit any additional contamination.
A microscopic examination revealed tiny critters that were moving quickly.
I suspect it might have been the frozen cell vial that was the contamination source.
Since I have been working with various cell lines, I have never seen anything like this!
Any suggestions as to what this might be, and how to get rid of it, would be of great help!
Hello Divya Singh
It would have been better if you had posted the picture of the contaminated culture. But since you have noticed turbidity in the media, it does indicate contamination. Since you have been working with various cell lines, I believe you have been maintaining sterility while working with cultures.
If only you could have posted the pic of the culture, I could have tried in the best possible way to identify the contaminant. Nevertheless, you could always streak a few microlitres of this so-called contaminated culture onto agar plate and incubate the plate in a 37-degree incubator overnight. The next day if you see colonies in the plate, it will confirm bacterial contamination.
You may be right that the frozen cell vial could be the contamination source. Without any delay you should discard the contaminated culture as other cultures in the incubator would be at risk. It is not advisable to treat the contaminated culture with a mixture of antibiotics (for bacterial contamination) unless it is a precious culture, and you cannot afford losing it. I would suggest that if you have more frozen vials of THP-1, you could thaw another vial but of a different lot.
Best.
@all Based on your description, it is likely that your THP-1 cell culture has been contaminated with a microbial organism. The presence of tiny critters moving quickly suggests a possible contamination by microorganisms, such as bacteria or protozoa. Here are some steps you can take to address the contamination issue:
If the contamination issue persists, it may be helpful to consult with a cell culture expert or the technical support team at your cell culture reagent supplier for further guidance and troubleshooting specific to THP-1 cells.
Remember, preventing contamination in cell culture requires attention to detail, proper aseptic techniques, and a clean working environment. Regularly reviewing and updating your cell culture protocols can help minimize the risk of contamination and ensure successful experiments.
- Badges
- Science topic
- Similar topics
- Phytochemistry
- Extracts