Hi,
I saw lots of cells debris in my cell culture. Lots of black spots which would not go even if I washed the cells. So, I performed DAPI staining and here is my result.
Can someone suggest if this is mycoplasma. What other tests can I do validate my observation?
I think the cells are contaminated with Mycoplasma.
You should use Plasmocin to kill the bacteria.
Possibly, i've had myco + cells that had similar DAPI staining pattern, but you should do a mycoplasma test to confirm which is quick. They're more common than people think.
https://www.thermofisher.com/tr/en/home/references/gibco-cell-culture-basics/biological-contamination/mycoplasma-contamination.html
What are other tests I can do to confirm my conclusions.
There was an anamoly that not all cells but very few had this staining result.
https://www.lonza.com/products-services/bio-research/cell-culture-products/mycoplasma-detection-and-removal/mycoalert-mycoplasma-detection-kit.aspx
https://www.lgcstandards-atcc.org/products/all/30-1012K.aspx?geo_country=tr
Hi ,
I wish you find these attached links useful for your problem solving.
https://bmcresnotes.biomedcentral.com/articles/10.1186/1756-0500-5-78
https://www.bionique.com/mycoplasma-resources/faq/mycoplasma-enter-cell-cultures.html
Best
Niloufar
Dear Pratik
- as said before I think your Hoescht staining indicates Mycoplama
- To unequivocally confirm mycoplama there are PCRs specific to Mycoplama rRNA: You take a small amount of your conditioned cell culture medium, perform PCR and then run on a 2% agarose gel as you would other rRNA products
- What is more the pcr test is more sensitive
- Pretty much all life sciences companies, Gibco, Thermofisher, GE health sell these Mycoplasma (rRNA) PCR kits
- There are ven more senstive colometric ELISA type kits but they are very expensive and with visible Nuclear spools derived from Mycoplasma cell infectioon he cheaper PCRs will be sensitive enough
- Badges
- Science topic