Problem of T Cell Proliferation Assay

Tom Thatcher ttha at uhura.cc.rochester.edu
Thu Mar 5 21:31:42 EST 1998


In article <6dml5s$c6g at ustsu10.ust.hk>,
CHAN Ella Wai Ching <boella at uxmail.ust.hk> wrote:
>Dear all,
>
>I am now planning to do a T cell proliferation assay to investigate whether
>the T cell from an injected mice can be stimulated to have strong 
>proliferation by  a protein (act as a stimulatin agent).

Immunize then harvest the draining lymph nodes.  The draining lymph nodes
can be used as is -- they contain T cells and accessory cells needed
for antigen presentation.  You can choose to purify the T cells.
If you do so you will need to add accessory cells back into the
culture.  The most common way to do this is to use irradiated spleen
cells (they must be irradiated to stop them from proliferating and
giving high background scores).  By far the easiest way to do the
experiment is to use the lymph node cells without additional purification.





>
>I would like to know whether it is necessary to purify the T cell suspension
>culture to remove the B cells and other accessory cells ???
>Can I just roughly purify the cell culture (by using nylon wool) and no 
>need to add extra accessory cells after purification. Coz' those accessory 
>cells are important for antigen presentation.  
>


-- 
Tom Thatcher                          | You can give a PC to a Homo habilis,
University of Rochester Cancer Center | and he'll use it, but he'll use it
ttha at uhura.cc.rochester.edu           | to crack nuts.



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