Hello
I have AML cells that have been cultured on stromal cells for weeks and would like to do flow staining looking for a specific antigen in the AML cells. I am not sure where these cells came from whether bone marrow or peripheral blood. In this case should I just collect the cells and stain or I do CD34 selection first to exlcude remaining stromal cells or others.?
Many thanks,
Abed
Hi Abdel Rahman.
Look if you area looking for AML Cells , either you have to sort them or make dual stain wz other LEUKEMIA specific antigens
Hi Abdulrahman,
You can take all of you cells and make a multicolor flow panel including CD34 and your antigen of interest using different fluorochromes for each. CD34 selection usually relies on magnetic bead separation, for which you need a relatively high number of cells; I am not sure you will have a high number of CD34 cells in your co-cultures and it doesn't seem the most efficient method to isolate your cells if you are planning to use flow afterwards.
If you are worried about contaminating non-hematopoietic cells you can also stain with CD45, which is a broadly used hematopoeitic surface marker.
Hope this helps!
Alba.
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