NK cell culture?

David Peritt Peritt_d at a1.mscf.upenn.edu
Thu Nov 9 08:56:53 EST 1995


In article <1995Nov3.113516.1 at muvms6> , lewis7 at muvms6.wvnet.edu writes:
>...i have been told that there is no possible way to culture NK cells on
any
>sort of medium, yet i do not believe that this is entirely true.  has
anyone
>had any recent luck in his/her research?

recipe for 8 day cultured and activated human NK cells:

.25x10*6 PBL/ml
.05x10*6 RPMI 8866 cells/ml
RPMI 1640 + 10% FCS

place in 24 well plates, 2 ml/well (yes a lot of plates and no flasks do
not work well.  Don't ask) We find 9% CO2 is a little better.
after 4 -5 days suck off majority of sup and add fresh media
let go another 3-4 days.
harvest cells

They are usually 70-80% NK and rest mosty CD8 T cells.  You can purify as
you wish.  We use rosetting or MACS beads.  Careful since NK are full of
FcR's



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