Hello,
I will do bone marrow transplantation in mice. For the moment I'm trying to deplete the BM from the donor of CD90.2 cells using MACS depletion (microbeads and magnetic column). However, at the end of the depletion I still have 29% CD90.2 positive cells in my 'negative' fraction.
Does somebody have experience with bone marrow depletion of T cells ??
Thank you a lot in advance
What about if you try monoclonal antibody (MoAb) (a complement fixing isotype MoAb) + complement (commercially available for that purpose from baby rabbit)...???
I think that the microbeads do not retain the cells very efficiently...!!! Probably the antibody on the beads does not bind the antigen on the desired cells with a high affinity...!
You can try immunodensity negative selection. This technique is less stressful for the cells and as sensitive as the MACS columns. There are several Rosettesep kits and you can design one.
https://www.stemcell.com/products/rosettesep-human-custom-cocktail.html
Pauline Janssen
Hi Pauline, have you solved this problem? If so, would you mind to share this effective BM T cell depletion protocol with me?
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