immunophenotyping - promyelocytes isolation (Jan/15/2006 )

Dear veterans,

I want to isolate promyelocytes from peripheral or bone marrow of acute promyelocytic leukemia patients. Please suggest whether CD 33 tagging alone will be sufficient. or should i use also CD 13, CD 11b etc., This is not my area, therefore bear with the mistakes if any. We have an automacs system. I would like to know the protocol and availability of the markers too.
Thank you.

Serena.

CD33 isolation is fine if you are not worried about having a pure APML isolation. APML cells are very hard to define by CD marker expression, the diagnosis of APML is normally based on morphology and cytochemistry. CD33 is a myeloid cell marker and so you will get some normal myeloid cells as well as the diseased cells. If this is the case then you could always do a PML protein stain and calculate the number of APML cells/total white cells to give you approximate purity. The best bet would be to do a double/triple isolation with the other CD markers you mentioned. Hope this helps!

Thank you for the reply and your suggestion on double/triple isolation. Iam only wondering about the viability of these cells and the stability of the RNA thro these procedure as we need to luk into expression profiles. I guess becoz of this, the selection of population shud be more stringent. please give your valuable suggestions and comments.
thank you once again,
serena