Lorenz R, Göllner T, Hempel K
Institute of Medical Radiation Research, University of Würzburg, Federal Republic of Germany.
Environ Health Perspect. 1990 Aug;88:129-32. doi: 10.1289/ehp.9088129.
Adult mice were Co-60 gamma irradiated, and 7 months later splenocytes were isolated, cultured in microwells, and the frequency of hprt-deficient mutants was determined by measuring the cloning efficiency in media with 6-thioguanine. The mutant frequency at 2, 4, and 6 Gy was 1.6 x 10(-5), 4.4 x 10(-5), and 12.7 x 10(-5), respectively. The frequency of spontaneous mutants was 2.5 x 10(-6). The effect of metabolic cooperation on the cloning efficiency of thioguanine-resistant T-cells in selective medium was evaluated in co-cultures with wild-type T-cells. We found that the growth of hprt-deficient T-cells is supported in the presence of thioguanine-inactivated wild-type splenocytes up to a cell density of 5 x 10(5) cells per well. When cell density was higher, cell growth was inhibited. Possibilities and limitations of cloned lymphocytes for the analysis of somatic mutations that occur in vivo are discussed.
成年小鼠接受钴 - 60γ射线照射，7个月后分离脾细胞，在微孔板中培养，通过测量在含6 - 硫鸟嘌呤的培养基中的克隆效率来确定hprt缺陷型突变体的频率。2 Gy、4 Gy和6 Gy剂量下的突变频率分别为1.6×10⁻⁵、4.4×10⁻⁵和12.7×10⁻⁵。自发突变体的频率为2.5×10⁻⁶。在与野生型T细胞共培养中评估了代谢合作对选择性培养基中硫鸟嘌呤抗性T细胞克隆效率的影响。我们发现，在硫鸟嘌呤失活的野生型脾细胞存在的情况下，hprt缺陷型T细胞的生长在每孔5×10⁵个细胞的细胞密度下得到支持。当细胞密度更高时，细胞生长受到抑制。讨论了克隆淋巴细胞用于分析体内发生的体细胞突变的可能性和局限性。
