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
