Munzer J S, Jones S K, O'Neill J P, Hartshorn J N, Robison S H
Department of Neurology, University of Vermont, College of Medicine, Burlington 05401.
Mutat Res. 1988 Sep;194(2):101-8. doi: 10.1016/0167-8817(88)90012-0.
The repair of DNA alkylation damage in human cells is poorly understood. We have adapted the alkaline elution technique for use with human peripheral blood lymphocytes in culture. We have also established conditions necessary for short-term culture of human lymphocytes. Lymphocyte growth which can be maintained for up to 30 days is dependent upon irradiated TK6 feeder cells and T-cell growth factor (crude TCGF). The amount of damage induced by a given concentration of methyl methane-sulfonate (MMS) is dependent upon cell number per ml of growth medium. The DNA damage measured, in lymphocytes, by alkaline elution is a composite of single strand breaks and alkali-labile lesions. Repair of this damage after appropriate recovery periods is also detectable. The irradiated feeder TK6 cells do not contribute to the number of strand breaks detected or the amount of recovery after treatment. This method offers a quick and reproducible means of detecting DNA damage and repair in human T-lymphocytes.
人类细胞中DNA烷基化损伤的修复机制尚不清楚。我们已将碱性洗脱技术应用于体外培养的人外周血淋巴细胞。我们还建立了人淋巴细胞短期培养所需的条件。淋巴细胞生长可维持长达30天，这取决于经辐照的TK6饲养细胞和T细胞生长因子（粗制TCGF）。给定浓度的甲磺酸甲酯（MMS）诱导的损伤量取决于每毫升生长培养基中的细胞数量。通过碱性洗脱测定的淋巴细胞中的DNA损伤是单链断裂和碱不稳定损伤的综合结果。在适当的恢复期后，这种损伤的修复也可被检测到。经辐照的饲养TK6细胞对检测到的链断裂数量或处理后的恢复量没有贡献。该方法为检测人类T淋巴细胞中的DNA损伤和修复提供了一种快速且可重复的手段。
