Reduced Macrophage viability

Hello,

I am working with Biogel-elicited macrophages and the last few months I have observed greatly reduced macrophage viability (even with 15h of culture, while before they were easily lasting for 3 days) and increased neutrophilia in my PECs. The viability of thioglycollate macrophages is not affected. And no obvious symptom is observed in my C57BL/6J.

I changed everything, including my Biogel Batch but I still observe macrophage death and neutrophilia (about 50% of my PECs!).  I am thinking it is a virus infection affecting specifically the biogel-type of macrophages as I can not classify all macrophage-elicitation methods as one.  But viruses like MHV affect both Biogel and thioglycollate. I don't want to swtich in the middle of my experiments to another cell type...   Any ideas about what could be causing this??

thank you!

Hi,

First, I would perform PCR on some samples from your mice to confirm it is MHV.  Alternatively, would it be possible for you to get some quarantined mice from a reputable supplier and attempt the isolation before they are in with the general mice stock and hence exposed to any circulating infection.

The paper below may help:
http://www.ncbi.nlm.nih.gov/pubmed/10024430

you can check if there is some problem with your culture media or FBS?