I have in the past collected at the end of 7 days ~50 million macrophages from 1 mouse.
I am in recent weeks getting about 10million macrophages from 1 mouse.
Please include precise nature of marrow extraction and disruption, plating dishes (adherent/non adherent), plating density, media and serum, additives, red blood cell lysis, and nature of MCSF and dose and media changes etc...
Working w/ bone marrow derived cells, I would suggest to run the BM through a ficool gradient, lyse the remaining RBCs, and plate them at 3x106/ml at least. They are "social cells" and therefore need closeness/confluence to be able to efficiently condition the media in their favor. I use 50 ng/mL of M/GM-CSF / mL. I work mostly with horse cells, and created protocols adapting from murine models. 10-20% horse serum or 10 FBS are fine. Horse serum seem to better preserve the HSC if that is your goal...
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