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 serum and freeze-killed feeder cells, has been developed for the mass culture of T-lymphocytes derived from human peripheral blood. In this protocol, bulk cultures can be cryopreserved approximately 8 days after initiation, and subsequent mass cultures generated a further week after recovery. At the end of this period, the lymphocytes have maintained a normal karyotype and cultures from different donors are very similar in terms of rate of cell division and expression of key antigenic markers. Background micronucleus frequencies and dose-responses for micronucleus induction by a reference clastogen, hycanthone, were also very similar in all the cultures examined. Such extended-term T-lymphocyte cultures are potentially valuable in genotoxicity testing, providing cells with the normal human karyotype which can be characterised and handled with the practical convenience of established rodent cell lines.
O'Donovan
M R
MR
Research Department, Boots Pharmaceuticals, Nottingham, UK.
Freemantle
M R
MR
Hull
G
G
Bell
D A
DA
Arlett
C F
CF
Cole
J
J
D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D002455:Cell Division; D002478:Cells, Cultured; D015925:Cryopreservation; D005615:Freezing; D005720:Gamma Rays; D005838:Genotype; D006801:Humans; D006826:Hycanthone / Q000633:toxicity; D007621:Karyotyping; D015162:Micronucleus Tests / Q000379:methods*; D016176:T-Lymphocyte Subsets / Q000187:drug effects / Q000276:immunology / Q000648:ultrastructure; D013601:T-Lymphocytes / Q000187:drug effects* / Q000276:immunology / Q000648:ultrastructure
D015703:Antigens, CD; D006826:Hycanthone