EXTENDED-TERM CULTURES OF HUMAN T-LYMPHOCYTES - A PRACTICAL ALTERNATIVE TO PRIMARY HUMAN-LYMPHOCYTES FOR USE IN GENOTOXICITY TESTING

A simplified method, using recombinant interleukin-2, foetal bovine se rum and freeze-killed feeder cells, has been developed for the mass cu lture of T-lymphocytes derived from human peripheral blood. In this pr otocol, bulk cultures can be cryopreserved similar to 8 days after ini tiation, and subsequent mass cultures generated a further week after r ecovery. At the end of this period, the lymphocytes have maintained a normal karyotype and cultures from different donors are very similar i n terms of rate of cell division and expression of key antigenic marke rs, Background micronucleus frequencies and dose-responses for micronu cleus induction by a reference clastogen, hycanthone, were also very s imilar in all the cultures examined. Such extended-term T-lymphocyte c ultures are potentially valuable in genotoxicity testing, providing ce lls with the normal human karyotype which can be characterised and han dled with the practical convenience of established rodent cell lines.