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Expansion of rat 6‐thioguanine‐resistant T‐lymphocyte clones by stimulation with ionomycin and a phorbol ester
Author(s) -
Chen Tao,
Aidoo Anane,
Casciano Daniel A.,
Heflich Robert H.
Publication year - 1998
Publication title -
environmental and molecular mutagenesis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1
H-Index - 87
eISSN - 1098-2280
pISSN - 0893-6692
DOI - 10.1002/(sici)1098-2280(1998)31:1<97::aid-em13>3.0.co;2-k
Subject(s) - ionomycin , clone (java method) , mutant , mutagen , lymphocyte , microbiology and biotechnology , biology , phorbol , mutation , stimulation , t lymphocyte , cell culture , ethyl methanesulfonate , genetics , biochemistry , dna , phosphorylation , endocrinology , in vitro , protein kinase c , gene
Previous molecular analyses of the mutations produced in the rat lymphocyte hprt assay were hindered by difficulties encountered in growing mutant lymphocytes from 6‐thioguanine‐resistant clones. In this study, we evaluated the ability of the calcium ionophore, ionomycin, and the tumor promotor, phorbol 12‐myristate 13‐acetate, to stimulate clone expansion. A medium containing these two agents, along with mitogen‐free conditioned medium, was found to expand 64% of 276 mutant clones to at least 5 × 10 5 cells in nine days of culture. Some clones were expanded to more than 4 × 10 6 cells. The procedure appears suitable for propagating rat lymphocyte clones for mutation analysis. Environ. Mol. Mutagen. 31:97–102, 1998. Published 1998 Wiley‐Liss, Inc. This article is a US Government work and, as such, is in the public domain in the United States of America.