Optimized culture systems for the preimplatation ICR mouse embryos with wide range of EDTA concentrations

In this study, in vitro preimplantation embryo culture media especially for outbred stock mice (Institute of Cancer Research (ICR)) were optimized with different concentrations of Ethylenediaminetetraacetic acid (EDTA). A plot with embryo development rates against EDTA concentrations ranging from 0 to 500µM showed a unique pattern with two characteristic peaks. 200µM was adopted as an optimal concentration of EDTA. The optimized media were also evaluated with two culture systems; conventional large volume culture system (1ml) and micro-droplet culture system. In the conventional large volume culture system, the blastocyst development rates were compared among three different media (F-10, KSOM and KSOM with the optimized 200µM EDTA). The rates were 0.4%, 16.7% and 57.6%, respectively. The development rates for micro-droplet (10µl) culture system was 73.9%. In conclusion, 200µM EDTA concentration in 10µl droplets in the KSOM medium was found as most suitable culture conditions for ICR mou...本研究针对远交系ICR小鼠（Institute of Cancer Research, ICR）优化了体外着床前胚胎培养液，通过调控乙二胺四乙酸（Ethylenediaminetetraacetic acid, EDTA）的不同浓度梯度完成配方优化。以0至500μM的EDTA浓度为横坐标、胚胎发育率为纵坐标绘制曲线，结果呈现出带有两个特征峰的独特分布模式，最终确定200μM为EDTA的最优添加浓度。随后，将优化后的培养液分别在两种培养体系中开展验证：常规大容量培养体系（1ml）与微滴培养体系。在常规大容量培养体系下，对比了F-10、KSOM以及添加200μM优化浓度EDTA的KSOM三种培养液的囊胚发育率，分别为0.4%、16.7%与57.6%；而10μl微滴培养体系的囊胚发育率可达73.9%。综上，在KSOM培养基中添加200μM EDTA并采用10μl微滴培养的体系，为ICR小鼠体外着床前胚胎培养的最优培养条件...