Hi all,

I have been doing BMDM cultures using L929 conditioned medium for quite some time. Recently few BMBDM batches have been behaving strangely. A population of mcarophages appears black and activated, while others appears round and healthy. Slowly by day 3 many such macrophages begin to appear and by the end of 7 hardly any macrophages appear normal.

I cleaned everything from medium to incubator after I had problems with the first BMDM batch, but the problem still persists. Has anyone faced this problem and found a solution to it? Any suggestions?

Many thanks
Aspee

Could this be mycoplasma contamination?

Tracy,

Not everything that goes wrong with cell culture has mycoplasma problem. Mycoplasma may take several days to infect, while they will not readily infect immuno-competant cells like macrophages.

The problem most likely is endotoxin, but I am not sure what is the source of it in my cultures.

Aspee