Can anybody please suggest me the specifc surface proteins (CDs) for asessing the granulocytic and macrophage differentiation of promyleocytic leukemic HL-60 cells?
CD11b on Granulocytes has higher expression than mono/Mac. Another feature of macrophage differentiation is enhanced granularity, (increases in side scatter (SSC) on flow cytometry).
Agree with Alex that you can distinguish granulocytes by higher SSC (granularity). Also, if you are characterizing mouse cells, for more accurate analysis you can assess expression of Ly6G and Ly-6C within CD11b+ cells. Granulocytes are positive for both markers (Ly-6G hi/Ly-6C low , while monocytes express Ly-6C exclusively.
@Alex and Rinat...Thanx for information....I agree with both of you...SSC is one of the choices to distinguish between the two populations ...but CD11b is expressed both by granulocytes and macrophages so doesn't seem to work perfectly here....I think there are specific CDs (surface markers) that can be used to differentiate between granulocytes and macrophages....Since I am using human leukemic HL-60 cells so Ly6G and Ly-6C expression will not work in my case....
I know you mentioned CD markers (others include CD16 and CD32-not sure how good they would stain in HL60) but maybe you could go 'old school' and look at NBT reducti on for Grans or monoyctic esterases. Morphology assessment (staining with Wrights-Giemsa).
Thanx Alex.....Indeed I am using one drug that was previously known to induce granulocytic differentiation but I have modified the culturing conditions and now I see that macrophages dominate granulocytes upon differentiation with the same drug. So I need to show the specific percentages of granuclocytes and macrophages in the same sample.....and I think in such a setting CDs seem to be the ultimate option....Any way thanx for your help....
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