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
