Hello all,
I am convinced that this is bacterial contamination of our Jurkat cell culture, though my team says it isn't. I lack the necessary experience. What do you think about this?
The Jurkat cells are incubated with RPMI1640, 10% FBS, 1%1xPen/Strep ; 5% CO2 ; 37°C ; humidified. Therefore, contamination is unlikely, but to my (unacquainted) eye I think so nevertheless.
Hello Gregor Böttiger It looks like contaminated flask. The reason can be handling error also. In next passage wash the cells with serum free media and also increase the FBS to 15%. Generally, jurkat cells look very clear round shape.
Gregor Böttiger is it possible to experiment in a separate flask with no antibiotics using the same cell line? If your original flask was contaminated this would lead to uninhibited bacterial growth which would be easier to visualize. Also, if your media is not its original color it is often another sign of bacterial growth, as your media contains a phenol red indicator.
There are many dying cells in your cell culture. I'm not sure though that it's due to bacterial infection (need higher magnification to confirm), more likely mycoplasma , or early yeast /algae contamination , or just nutrients deprivation. You can add antibiotic-antimycotic (instead of p/s) or normacin (have anti-mycoplasma activity) into culture, Ficoll it to remove dead cells, wash cells well and start a new culture . Some people add 10mM HEPES into Jurkat cell culture medium.
I have experience culturing Jurkat cells and this looks like a bacterial infection. Sometimes cell death and cell debris can look somewhat similar. Observe carefully under high magnification if these structures show fine twitching movement, then for sure its infection. You can also take this media and split it into two flask/dish and treat one with antibiotics and leave the other as it is. (Careful though that can create contamination in other flasks). You can keep the flasks tightly closed in an incubator for 12 hrs and you will see the difference. Best
Hi there!
I`ve worked with Jurkats alittle (now don`t work). In my memory they always looked strange and although they were fine, I was always scared that they are dying. Your pictures look even worse... Looks like many cells are dying...
You can remove antibiotics, observe for some time and test for mycoplasma. If bacteria are there, they will grow very fast. If it is mycoplasma, PCR must detect it even in culture medium.
One more option is suboptimal culture conditions. Consider addition of the glutamine and/or b-ME and/or pyruvate. And remove antibiotics.
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