The culture of preimplantation embryos in vitro is an important method for human and mouse reproductive technology. This study aims to investigate the influence of different conditions of culture media on the preimplantation stage of mouse embryos cultured in vitro, and monitor the post-implantation development of new mice after embryo transfer to surrogate females. We demonstrated here that mouse embryos cultured in vitro in fresh M16, KSOM, Global, and HTF embryo culture media from one cell to the blastocyst stage and the subsequent embryo transfer to surrogate females are able to proceed through post-implantation development and, after birth, develop into healthy mice. However, culture of embryos in differently aged media shows various (often unpredictable) results. To find the optimal storage conditions of culture media, we suggest that the freezing and long-term storage of these media at − 80°C will not influence the quality of the media. To test this hypothesis, we grew embryos from one cell to blastocysts in vitro in the selected media after thawing and subsequently transferring them to surrogate females. Embryo culture in these four media after thawing does not affect preimplantation and postnatal mouse development. Thus, we have shown that storage of embryo culture media at low temperature (− 80°C) does not impact the quality of the media, and subsequently, it can be used for the culture of embryos for the full preimplantation period, the same as in fresh media.

植入前胚胎体外培养是人类和小鼠生殖技术的重要方法。本研究旨在探讨不同培养基条件对体外培养小鼠胚胎着床前阶段的影响，并监测胚胎移植至代孕雌性后新小鼠的着床后发育情况。我们在此证明，在新鲜的 M16、KSOM、Global 和 HTF 胚胎培养基中体外培养的小鼠胚胎从一个细胞到胚泡阶段以及随后的胚胎移植到代孕雌性体内能够进行植入后发育，并在出生后进行，发育成健康小鼠。然而，在不同老化介质中培养胚胎会显示出不同的（通常是不可预测的）结果。为了找到培养基的最佳储存条件，我们建议将这些培养基冷冻并在-80°C长期储存不会影响培养基的质量。为了检验这一假设，我们在解冻后在选定的培养基中在体外将胚胎从一个细胞培育成囊胚，然后将它们转移给代孕雌性。解冻后在这四种培养基中进行胚胎培养不会影响植入前和出生后小鼠的发育。因此，我们已经证明，胚胎培养基在低温（− 80°C）下储存不会影响培养基的质量，随后，它可以用于整个植入前阶段的胚胎培养，与在新鲜媒体中。