Cloning CTL's in New Species - How?
MMAGEE at TCID.TDH.STATE.TX.US
Mon Apr 8 07:42:21 EST 1996
> So, how would I go about cloning and propagating antigen-specific
> CTL's? In the bad old days before Il-2, did people use irradiated feeder
> cells? Would PBMC feeders suffice? Sacrificing an animal and isolating
> splenocytes would be out of the question for my experiment. I want to
> accomplish this with bleeding only.
You could take peripheral blood and stimulate it with Concanavalin A
(I used 5 micrograms Con A/ml with a cell concentration of 5 x
10^6/ml). After 48 hours, spin the culture to remove cells, add
alpha-methyl mannoside to bind residual Con A, and filter through a
0.22 micron filter. Use this conditioned media at a 10% final
concentration with your cultured cells. I have had very good luck
with this source of growth factors.
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Email: MMAGEE at tcid.tdh.state.tx.us
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