Since I want to perform experiments at different time points, I would like to culture MDSCs so that they can be used after 48 hours of being isolated from the patients' blood. I want to be able to co-culture MDSCs with the T cells from the same patient but only after 48 hours. Does anyone have experience with this?
Hi,
I advise you to check out a Miltneyi positive-selection magnetic column separation loaded with CD33 beads.
https://www.miltenyibiotec.com/US-en/products/macs-cell-separation/cell-separation-reagents/microbeads-and-isolation-kits/granulocytes-and-myeloid-cells/cd33-microbeads-human.html
As for using T-cell suppression to define MDSC.I believe that what you will isolate will provide a negative control If you isolate from the PB of a cancer patient you will have much more success.
Ps; MDSCs and their next of kin CFU-GM or myeloid precursors require growth factors to avoid apoptosis. Thus, in the absence of G-CSF, GM-CSF, M-CSF, or SCF they will apoptosis within hours.
Walid Hassene Hamri thanks for your kind answer.
The protocol for isolating the two subsets of MDSCs with beads is well established in my group. The question I now have is more about how to keep these cells in culture. According to what you explain to me I must keep them with growth factors. Maybe you know of a study that you can recommend where I can read about how they culture them? Thanks so much in advance.
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