Help: Growing Human T4 Helper Cells

[USER]

RRRP87C at prodigy.com (Matthew Dubeck) wrote:
>I would like to grow human T4 helper cells in some sort of growth medium, 
>but need some direction as to where I should start.  Can anyone send me a 
>reference to a procedure for maintaining a human T4 helper culture?  
>Thanks.
>
>                                                Matthew Dubeck
>                                                RRRP87C at prodigy.com
>


In one 24 well-plate put per well in one ml:

200,000 CD4 cells
one million irradiated (30 Gy) PBL from different donors
100,000 irradiated (50 Gy) EBV-B cells

in one ml of medium (Iscove's or RPMI)
with 5-10 % of (preferably) human serum, but FCS will work too.
0.5-1 ug/ml PHA
10-50 u/ml rIL-2 (Cetus units)

Split growing wells and refresh medium with:

medium + serum + IL2


Good luck,

If you need more help, mail me at:

d.orsini at elsevier.nl


Daniela