Suyama I, Etoh H
Mutat Res. 1983 Jan;107(1):111-8. doi: 10.1016/0027-5107(83)90082-9.
A microculture technique was applied to the study of lymphocytes of Umbra limi, which have a low number of large meta- and submetacentric chromosomes (2n = 22). On the 5th day (90h) and later after initiation of culture at 20 degrees C, some cultures provided well spread metaphase chromosomes for analyses. After initiation, cultures were irradiated with 50, 100, 150 and 200 R of 200 kVp X-rays. The cultures were harvested on the 5th day, at which time all arrested metaphase chromosomes were in their first division. The dicentric yields induced in X-irradiated Umbra lymphocytes were observed to be significantly (P = 0.05) lower than those in human lymphocytes. The resulting dose-response relationship for dicentric yield was described by the quadratic equation Y = aD + bD2. The advantage of this method lies in the fact that small amounts (0.1 ml) of blood can be repeatedly withdrawn from the fish after a minimal interval of 2 weeks. The microculture technique with Umbra lymphocytes is thought to have a wide range of applications in studies of the potential genotoxic effects of clastogenic agents in the aquatic environment.
一种微量培养技术被应用于溪鳉淋巴细胞的研究，溪鳉具有数量较少的大型中着丝粒和亚中着丝粒染色体（2n = 22）。在20℃开始培养后的第5天（90小时）及之后，一些培养物提供了良好伸展的中期染色体用于分析。培养开始后，用200 kVp的X射线分别以50、100、150和200伦琴进行照射。在第5天收获培养物，此时所有停滞在中期的染色体都处于第一次分裂阶段。观察到X射线照射的溪鳉淋巴细胞中诱导产生的双着丝粒产率显著低于人类淋巴细胞（P = 0.05）。双着丝粒产率的剂量 - 反应关系用二次方程Y = aD + bD²来描述。该方法的优点在于，在最短间隔2周后，可以从鱼身上反复抽取少量（0.1毫升）血液。溪鳉淋巴细胞的微量培养技术被认为在研究水生环境中致断裂剂的潜在遗传毒性效应方面具有广泛的应用。
