Porter A J, Singh S M
Department of Zoology, Children's Hospital of Western Ontario, University of Western Ontario, London, Canada.
Mutagenesis. 1988 Jan;3(1):45-9. doi: 10.1093/mutage/3.1.45.
We have worked out a simplified method to detect embryotoxicity associated with blood serum. Here we report results for serum from female rats treated with 0, 25, 250 and 500 mg/kg cyclophosphamide (CP) as a control experiment in order to evaluate the feasibility of this protocol. Eight-cell mouse embryos were cultured for 48 h in medium supplemented with 10% rat serum and 10(-6) M 5-bromodeoxyuridine (BrdU). Colchicine (0.2 microM) was added 3 h prior to harvesting the cells for chromosome preparation, and differentially stained sister chromatids were visualized after fluorescence-plus-Giemsa staining. Rats treated with increasing doses of CP yielded serum which had increasing mutagenic [as evaluated by sister-chromatid exchange (SCE)] and toxic effects on mouse embryos during 48-h in vitro cultures. The SCEs/cell (based on 40 chromosomes) ranged from 17.6 +/- 0.31 (mean +/- SE) for serum from control (0 mg/kg CP) rats to 40.4 +/- 0.22 for serum from rats given 250 mg/kg CP. Serum from rats treated with 500 mg/kg CP was essentially lethal on mouse embryos. The percentage embryo survival decreased from 79.8 +/- 0.31% (mean +/- SE) for serum from control rats to 8.4 +/- 0.42% for serum from rats given 500 mg/kg CP. These results indicate that after appropriate testing, this protocol could be used for the possible detection of embryo genotoxicity associated with any human serum. If successful, the evaluation of mouse embryo survival and SCE in the presence of human female sera could become an 'early warning' system for possible reproductive problems (e.g. recurrent abortions, low birth weight and malformations) of unknown aetiologies.(ABSTRACT TRUNCATED AT 250 WORDS)
我们研发出一种用于检测与血清相关的胚胎毒性的简化方法。在此，我们报告以0、25、250和500mg/kg环磷酰胺（CP）处理的雌性大鼠血清的检测结果，作为对照实验，以评估该方案的可行性。将八细胞期小鼠胚胎在补充有10%大鼠血清和10(-6)M 5-溴脱氧尿苷（BrdU）的培养基中培养48小时。在收获细胞用于染色体制备前3小时加入秋水仙碱（0.2 microM），荧光加吉姆萨染色后可观察到差异染色的姐妹染色单体。给予递增剂量CP处理的大鼠所产生的血清，在48小时体外培养期间，对小鼠胚胎具有递增的诱变作用[通过姐妹染色单体交换（SCE）评估]和毒性作用。每个细胞的SCEs（基于40条染色体）范围从对照（0mg/kg CP）大鼠血清的17.6±0.31（平均值±标准误）到给予250mg/kg CP大鼠血清的40.4±0.22。给予500mg/kg CP处理的大鼠血清对小鼠胚胎基本具有致死性。胚胎存活率百分比从对照大鼠血清的79.8±0.31%（平均值±标准误）降至给予500mg/kg CP大鼠血清的8.4±0.42%。这些结果表明，经过适当测试后，该方案可用于检测与任何人类血清相关的胚胎遗传毒性。如果成功，在存在人类女性血清的情况下评估小鼠胚胎存活率和SCE，可能成为针对不明病因的潜在生殖问题（如习惯性流产、低出生体重和畸形）的“早期预警”系统。（摘要截选至250字）
