I would like to isolate NK from PBMC by positive magnetic selection to obtain an untouched, NK-less PBMC fraction. The only commercial kits with positive NK selection I came across, used a preceding enrichment step consisting of a negative NK selection. In case I missed one, does anybody know a kit which only applies positive magnetic NK isolation? Buffy coats are used as starting material.
Dear Jorrit de Waele,
why is it principal to use positive selection kit? If you need other non-NK cells, they will be on column and you can pull them with plunger.
Probably you need to obtain particles - free PBMC? The only solution there is to use anti CD56 antibodies, but positive fraction will contain at least NKT cells too. May be the next step is to use NKT cell positive isolation kit?
Dear Vera
Indeed, I would prefer to obtain particles-free PBMC in order for them to stay untouched. The NK cells can be touched, since I want to investigate the PBMC without these NK cells. The solution you provide was the only other one I could come up with as wel, but as you mention, at least NKT cells will be selected next to NK cells as well.
Thanks for your opinion!
Hello Jorrit,
As Vera says, if you do a CD56 positive selection, you'll get a mixed population, but at least you'll have your CD56- population untouched and ready to go. I work at Stemcell Tech. and would recommend the EasySep™ Human CD56 Positive Selection Kit . The negative fraction will be very easy to get and won't involve trying to plunge cells out of a column.
Dear Cyrus
Thank you for the information, this might be an elegant solution. I have requested a demo at StemCell Tech.
I used the NK isolation kit both the CD56 positive and negative selection with magnetic beads from Mylteny. With the CD56 positive selection you will loose less NK cell. However in my case, (in the uterous) this kit detects only NK cells and I don't need to bother too much about NKT. In the blood I would rather prefer to do a CD3 negative selection (Mylteny) of the CD56 purified cells. You can try first checking the CD56 enrichment by FACS and if you need an higher purity you can try the CD3 kit. I am saying this, because it has been recently debated how sometimes enrichments can introduce huge problems in case you are not checking them carefully (see the publication I've linked)
http://www.ncbi.nlm.nih.gov/pubmed/24337374
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