Extended-term cultures of human T-lymphocytes: a practical alternative to primary human lymphocytes for use in genotoxicity testing

• Published in: Mutagenesis Vol. 10; no. 3; p. 189
• Main Authors: O'Donovan, M R, Freemantle, M R, Hull, G, Bell, D A, Arlett, C F, Cole, J
• Format: Journal Article
• Language: English
• Published: England 01.05.1995
• Subjects: Animals; Antigens, CD - metabolism; Cell Division; Cells, Cultured; Cryopreservation; Freezing; Gamma Rays; Genotype; Humans; Hycanthone - toxicity; Karyotyping; Micronucleus Tests - methods; T-Lymphocyte Subsets - drug effects; T-Lymphocyte Subsets - immunology; T-Lymphocyte Subsets - ultrastructure; T-Lymphocytes - drug effects; T-Lymphocytes - immunology; T-Lymphocytes - ultrastructure
• ISSN: 0267-8357
• DOI: 10.1093/mutage/10.3.189

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.