A modified protocol for the cytochalasin B in vitro micronucleus assay using whole human blood or separated lymphocyte cultures.
A modified protocol for the cytochalasin B in vitro micronucleus assay using whole human blood or separated lymphocyte cultures.
复制标题
使用全人血或分离的淋巴细胞培养物进行细胞松弛素 B 体外微核测定的修改方案。
作者：
S. Ellard;E. Parry
描述了全血或分离淋巴细胞培养物中微核体外分析的改良方案。在两个收获时间（68或72小时）检查了不同浓度的细胞松弛素B（3、4.5或6微克/ml）对双核细胞的诱导。通过在起始后44 h向培养物中加入6 μ g/ml剂量的细胞松弛素B，并在24 h（全血）或28 h（分离的淋巴细胞）后收获，获得了最佳产量。用吖啶橙子对淋巴细胞的细胞离心制备物（在试验开始时或收获时使用Ficoll商业制备物从全血中分离）进行染色。使用这种方法，微核阻断的淋巴细胞保持完整，并且很容易识别微核。该方法适用于全血和分离的淋巴细胞培养物，因此可以直接比较对遗传毒性试剂的敏感性。
A modified protocol is described for the in vitro analysis of micronuclei in whole blood or separated lymphocyte cultures. The induction of binucleate cells by various concentrations of cytochalasin B (3, 4.5 or 6 micrograms/ml) was examined at two harvest times (68 or 72 h). An optimal yield was obtained by adding cytochalasin B at a dose of 6 micrograms/ml to cultures 44 h after initiation with harvest 24 h (whole blood) or 28 h (separated lymphocytes) later. Cytocentrifuge preparations of lymphocytes (separated from whole blood using commercial preparations of Ficoll either at the commencement of the assay or upon harvest) were stained with Acridine Orange. Using this method, cytokinesis-blocked lymphocytes remain intact and micronuclei are readily identified. The method is suitable for both whole blood and separated lymphocyte cultures, thus allowing direct comparisons of sensitivity to genotoxic agents.