I am trying to develop macrophages from BALB/c bone marrow with an isolated polysaccharide of my own interest. During experiment i observed 45- 50% F4/80+ cells by FACS in 7 day culture (total non adherent and adherent fraction taken)of bone marrow (simple bone marrow culture/ adding nothing; No MCSF and No polysaccharide of my interest). Cells were detected CD14-. I am little bit confused as i found 2-5% in some references. Please suggest me the logical percentage of F4/80+ cells in SIMPLE 7 day culture of bone marrow (media- RPMI+ 10% FCS).
You are making the assumptions that the bone marrow from all mice are equal, unless pooled into a homgenous population, an unwise assumption. Also, you are making the assumption that your 10% FCS does not contain bioactive factors that might alter the results of your experiments. Also a bad assumption. Suggest pooling bone marrow to derived homologous population and use heat inactivated serum.
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