An improved procedure for the in vitro expansion of human T-lymphocyte clones for mutant analysis.
Assays detecting mutants present in human peripheral blood T-lymphocytes have been developed to monitor the population for somatic mutations. In order to investigate the nature of the mutations, colonies are further expanded in vitro by repeated lectin stimulation. To characterise fully each mutant clone, sufficient cells (approximately 10(7)) must be available for several molecular and biochemical techniques to be employed. These techniques, and their importance to the assay for population monitoring, are discussed briefly. We report here that the expansion of mutant colonies to approximately 10(7) cells by repeated lectin stimulation is not effective for all T-cell clones but that an alternative "lectin free" expansion method has enabled us to expand all the clones tested from a variety of normal donors and other individuals of interest.
Beare
D M
DM
MRC Cell Mutation Unit, University of Sussex, Falmer, Brighton, UK.
Aldridge
K E
KE
O'Donovan
M R
MR
Cole
J
J
D002455:Cell Division / Q000187:drug effects; D002469:Cell Separation; D002999:Clone Cells; D006801:Humans; D007376:Interleukin-2 / Q000494:pharmacology; D009154:Mutation; D010835:Phytohemagglutinins / Q000494:pharmacology; D015375:Receptors, Interleukin-2 / Q000378:metabolism; D013601:T-Lymphocytes / Q000166:cytology* / Q000187:drug effects
D007376:Interleukin-2; D010835:Phytohemagglutinins; D015375:Receptors, Interleukin-2