College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, 712100, China; and Key Laboratory of Animal Biotechnology, Ministry of Agriculture, Yangling, Shaanxi, 712100, China.
College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, 712100, China; and Key Laboratory of Animal Biotechnology, Ministry of Agriculture, Yangling, Shaanxi, 712100, China; and Corresponding author: Email:
Reprod Fertil Dev. 2020 Jun;32(9):879-884. doi: 10.1071/RD19086.
This study investigated the possibility of a sealed culture system in polymerase chain reaction (PCR) tubes to maintain embryo development. The embryo density that could support the development of 2-cell stage mouse embryos to the hatching stage was determined. At an embryo density of 1:2 (100 embryos cultured in 200μL CZB medium that had been pretreated with a reference gas containing 5% O2), the developmental rate was higher and fewer embryos exhibited reactive oxygen species- or hypoxia-induced injury compared with other sealed culture groups. Expression of growth factors (insulin-like growth factor (IGF) 1, IGF2, epidermal growth factor and transforming growth factor-α) and their receptors was evaluated, with similar expression patterns seen for embryos in sealed culture (5% O2, embryo density of 1:2) compared with the control group (embryos cultured in microdrops and placed in a 37°C, 5% CO2 water-jacketed incubator; P>0.05). After transfer of blastocysts generated by the sealed culture into recipients, there were no obvious differences in the rate of normal live pups births between the sealed culture and control groups (P>0.05). Thus, the sealed embryo culture system in PCR tubes is feasible for use in situations which cannot use a traditional incubator, such as in space and during the transport of embryos.
本研究探讨了在聚合酶链反应（PCR）管中密封培养系统以维持胚胎发育的可能性。确定了支持 2 细胞期小鼠胚胎发育至孵化阶段的胚胎密度。在胚胎密度为 1:2（在经过参考气体预处理的 200μL CZB 培养基中培养 100 个胚胎，参考气体中含有 5%O2）时，与其他密封培养组相比，发育率更高，且较少的胚胎表现出活性氧或缺氧诱导的损伤。评估了生长因子（胰岛素样生长因子（IGF）1、IGF2、表皮生长因子和转化生长因子-α）及其受体的表达，与对照组（在微滴中培养的胚胎，并放置在 37°C、5%CO2 水浴孵育箱中）相比，密封培养（5%O2，胚胎密度为 1:2）中的胚胎具有相似的表达模式（P>0.05）。将通过密封培养产生的囊胚移植到受体中后，密封培养组和对照组之间的正常活产仔率没有明显差异（P>0.05）。因此，PCR 管中的密封胚胎培养系统可用于传统孵育器无法使用的情况，例如在太空和胚胎运输期间。
