Hi Tony:
You could always try injecting some india ink or some other
inert particulate (e.g. coloured latex microbeads or the fluorescent ones
used to calibrate FACS machines, or bacteria) into the peritoneal cavities
of the deer mice a few hours before performing your lavage. Examining
smears or cytospins of the lavaged cells will reveal whether they contain
monocytes (they should have ingested the inoculated particulates) which for
whatever reason fail to adhere to glass/plastic? N.B. even without
injecting an inflammogen there should still be a sizeable population of
resident peritoneal macrophages, if they resemble lab mice that is.
Finally, you could experiment with other inflammogens (e.g. proteose
peptone, sodium caseinate) which might cause recruitment of greater numbers
of monocytes. Incidentally, were there any granulocytes in the lavages
from the deer mice?
Regards,
Wayne.
Tony Schountz wrote:
> I'm having a bit of a problem that I hope someone might be able to help
> me with. I'm trying to harvest peritoneal macrophages from deer mice,
> but the cells that I get do not look like macrophages because they do
> not adhere to tissue culture plates. My protocol is to inject deer mice
> and BALB/c with 1.5 ml of 3% thioglycolate then harvest 3-5 days later.
> The BALB/c clearly give us macrophages, but the deer mice do not. Any
> suggestions for alternative macrophage harvesting?
>> Thanks,
>> Tony
>> --
> Tony Schountz, Ph.D.
> Department of Biological Sciences
> Mesa State College
> mailto:tschount at mesastate.edu