I am working with lymphocyte cultures for micronucleus test. I am using PHA-M in my assays. I have a curiosity to use Concanavalin A in place of PHA-M to stimulate the lymphocytes. Can anyone suggest me the pros and cons while using concanavalin in the cultures?
3 questions that might help you get more accurate information:
1) Are you working with total lymphocytes or is there a particular population of lymphocytes that you care most about?
2) What is your read-out for stimulation (e.g. proliferation, CD69 upregulation) and how long after stimulation are you harvesting the cells?
3) What species are you working with?
I have never tried PHA-M, but I have done a lot with ConcanavalinA. It works well at 2ug/ml for inducing stimulation human T cells to proliferate. However, using mouse cells, CD8 T cells proliferate well, but CD4 T cells don't after ConA.
If you are working with T cells, there are more physiologic ways to stimulate them, such as using anti-CD3 (and anti-CD28 if you want costimulation).
ConA and PHA-M are lectins wich recognize different moieties in cell surface but in general terms, produce similar mitogenic stimuli in PBMC. There are subpopulations that are more responsive to conA (such as some T cells and B cells) than to PHA. I believe that for your purpose (clastogenesis study) they can be interchangeably used. Be aware that PHA is unstable in the fridge in the way that its activity may decay from week to week. So, it is advisable to stock concentrated aliquots and dilute them at the moment of use. Good luck!
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