Kanda Reiko, Shang Yi, Tsuji Satsuki, Eguchi-Kasai Kiyomi, Hayata Isamu
Radiation Hazards Research Group, National Institute of Radiological Sciences, 4-9-1, Anagawa, 263-8555, Inage-ku, Chiba, Japan.
Biosci Rep. 2004 Dec;24(6):641-50. doi: 10.1007/s10540-005-2798-4.
There is an incentive to develop a culture system of mouse peripheral blood lymphocytes (PBLs) to serve as models for studying genotoxic effects in humans exposed to mutagens, including ionizing radiation. However, many past approaches have been laborious, complex and only partly reproducible. In the present study, we established an improved culture system of mouse PBLs by removing blood and/or plasma, which was found to inhibit in vitro mitotic stimulation or proceeding cell cycles of lymphocytes. We compared the reactions of isolated PBLs to mitogens between the classical method and the present improved one. Then, we applied this method to the cytogenetic analysis using chemically induced premature chromosome condensation (PCC) as well as the conventional analysis, and demonstrated that the frequency of excess fragments observed in PCC cells might be useful to quantify the radiation-induced damages on chromosomes.
开发一种小鼠外周血淋巴细胞（PBLs）培养系统作为研究暴露于诱变剂（包括电离辐射）的人类遗传毒性效应模型具有一定的激励作用。然而，过去的许多方法都很费力、复杂且仅部分可重复。在本研究中，我们通过去除血液和/或血浆建立了一种改进的小鼠PBLs培养系统，发现血液和/或血浆会抑制淋巴细胞的体外有丝分裂刺激或细胞周期进程。我们比较了经典方法和本改进方法中分离的PBLs对有丝分裂原的反应。然后，我们将该方法应用于化学诱导早熟染色体凝聚（PCC）的细胞遗传学分析以及传统分析，并证明在PCC细胞中观察到的过量片段频率可能有助于量化辐射对染色体的损伤。
