T cell proliferation assay

Kang Liu kang.liu at worldnet.att.net
Thu Sep 23 19:01:55 EST 1999

I am doing TCR transgenic T cell proliferation assay in vitro. (OT-I CD8 T
cells and DO.11 CD4 T cells). There are three questions:

1. For 96-well plate, how many T cell should be used in each well? I used
200,000 cells/well, is that too high? And after how many days coculture with
APC should H3 thymidine be added?

2. Which medium is good for the assay? I only used RPMI, any one has better
experience with other medium?

3. Does it make difference to use mouse serum v.s. FCS? What percentage of
serum should be added to the medium?

Thanks for help.

e-mail: Liuk at rockvax.rockefeller.edu

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