Generate DC from human PBMC - (Oct/10/2005 )
Hello, I am generating mature DC from human PBMC. I have a couple of protocols, I am wondering the amount of GM-CSF and IL-4? and the positive percentage of the mature DC is only around 3%. Anybody would like to share your experience here or paste your protocol?
Thank you very much
Sorry, the DC positive percentage is around 33%, not 3%.
I was told that it can be reached 100%. My question is how can I generate more DC and higher percentage of mature DC from human PBMC?
we use the standard protocal of 100ng/ml of GM-CSF and 20ng/ml of IL-4 though you can go down to 25ng/ml of GM-CSF and still be fine. The only way you can generally reach a 100% maturation is to use a strong stimulus like LPS or Calcium ionophore. If you are not using that you will most likely never see 100% at 24 hrs. DCs are a maturing heterogenous population so its pretty hard to get that 100%. Hope this helps
Thank you for your message. I will try.
Have a great weekend.