ABSTRACT
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.
Subject(s)
Micronucleus Tests/methods , T-Lymphocytes/drug effects , Animals , Antigens, CD/metabolism , Cell Division , Cells, Cultured , Cryopreservation , Freezing , Gamma Rays , Genotype , Humans , Hycanthone/toxicity , Karyotyping , T-Lymphocyte Subsets/drug effects , T-Lymphocyte Subsets/immunology , T-Lymphocyte Subsets/ultrastructure , T-Lymphocytes/immunology , T-Lymphocytes/ultrastructureABSTRACT
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, aliquots of bulk cultures can be cryopreserved about 8 days after initiation, and subsequent mass cultures generated a further week after recovery. At the end of this period the lymphocytes are karyotypically normal, and cultures from different donors are very similar in terms of rate of cell division and expression of key antigenic markers. Such extended-term T lymphocyte cultures are potentially valuable in genotoxicity testing, combining the practical convenience of established cell lines with the theoretical advantage of possession of the normal human karyotype.