Sick primary cells - (Dec/02/2005 )
We have been battling low viablities in our dendritic cell cultures for a while now. Attached are two pictures of our super sad cells:
We think that maybe the little structures are responsible for our sick looking cells (that or a consequence of something else).
We have tried almost everything from changing cytokines like IL-4 and GM-CSF to different media etc.
Anyone know what might be going on here?
Any help would be great!!!
I haven't worked with primary dendritic cells but have done primary culture work with epithelial cells. Some thoughts:
Has this happened only with recent cultures? A change in the lot# of serum used in media can make a difference as with any change in a reagent you use for isolating and growing the cells. If you have changed something recently, you may need try to order from the original lot# or try other lot#s if you're unable to do so.
Any possibility of mycoplasma infection? This can cause poor growth. You can check for mycoplasma with a detection kit.
Is the incubator working properly? You may want to check the CO2 level and make sure the sensor is working properly.
You could also try adding additional glutamine to the media. Glutamine is degraded fairly quickly even in unopened media so I always add extra.