Trouble obtaining T cell clones!
DARREN at PARASIT.Lan.McGill.CA
Mon Oct 2 14:15:18 EST 1995
I have been having limited success obtaining murine T cell clones
against my antigen of interest. I easily get T cell lines
isolated from the inguinal nodes of Balb/c mice immunised
subcutaneously with antigen emulsified in complete Freund's adjuvant.
However it is proving difficult to establish T cell clones from these
lines (passaged every 14 days in the presence of antigen and IL-
2) even though their proliferative responses to antigen are quite
I clone the cells in 96 well plates in the presence of 1 million
mitomycin-C treated spleen cells, 50 ug/ml antigen and 10 U/ml IL-2.
I have varied the numbers of T cells/well from 10 to 500, however, I
either get no clones or too many (suggesting that the positive wells
are not monoclonal).
As there are not many detailed T cell cloning protocol around, I'd
appreciate any comments, suggestions or tips that anyone has. Many
Cloneless in Montreal
J. Darren Campbell
Institute of Parasitology,
Macdonald Campus of McGill University
21,111 Lakeshore Road, Ste-Anne-de-Bellevue,
Quebec, Canada H9X 3V9
e mail: Darren at Parasit.lan.mcgill.ca
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