I am culturing primary human brain stem cells which grow as neurospheres but I seem to constantly get what looks like bacterial contaminations and cannot seem to work out why. I have had 4 years cell culture experience with 1 contamination up to this point and maintain stringent asceptic techniques so I am very unsure why this keeps happening.
Any help would be appreciated!
Can you upload a photo of the culture or define how it looks like???
best wishes
Hi Sophie,
Have you also been culturing these primary cells for a while? Did you get the cells from a new source that maybe causing the contamination?
-ED
This is what the cells look like. They grow very well and grow in neurospheres as you can see. The media does not smell and remains clear. I have been culturing this specific line for 2 months. I have also tested the media bu adding 100ul to 1ml LB broth and the media did not change to cloudy over 48hrs.
Hi Sophie,
Are your suspected bacteria in the picture you posted in this thread? If they are not cloudy in your culture media or LB I think you should'nt worry a lot unless your results just does not make sense. I would guess the grainy particles in your culture could be cell debris from cells that did not survive culturing. Can you see if the debris comes later in your culture rather than the day after your start?
-ED
The debris comes almost always the day after culture. I think you may be right that it is cell debris. I will try to be more gentle with the cells during culture and see if I can reduce it. Thankyou for your help!
As emmanuel said, I am pretty sure that they are cell debris. In addition, congratulations for such nice neurospheres.
Best wishes!!!
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