Conaghan J, Handyside A H, Winston R M, Leese H J
Institute of Obstetrics and Gynaecology, Royal Postgraduate Medical School, Hammersmith Hospital, London, UK.
J Reprod Fertil. 1993 Sep;99(1):87-95. doi: 10.1530/jrf.0.0990087.
Although human embryos will develop in vitro for six days or more, little is known about the effects of the primary nutrients, pyruvate and glucose, on development. Because the nutrient requirements of embryos change throughout preimplantation development, the effects of altering substrate concentrations in the culture medium were examined, using 'surplus' human preimplantation embryos cultured from the two-four-cell stage to the blastocyst stage in medium containing various concentrations of pyruvate and glucose. Between the one-cell stage and the two-four-cell stage all of the embryos were exposed to 0.47 mmol pyruvate l-1 and 5.5 mmol glucose l-1. Pyruvate as sole substrate in the medium could support blastocyst development to an extent of 59% (10 of 17). Conversely, culture of embryos in pyruvate-free medium resulted in the developmental arrest of 84% (21 of 25) of embryos, and for the 16% (4 of 25) that did reach the blastocyst stage there was a significant decrease in metabolic activity on day 4-5, during the morula to blastocyst stage transition. Embryos could not use glucose to compensate for the lack of pyruvate in the medium. Pyruvate uptake was related to exogenous concentration and optimal development occurred at the highest concentration tested, 0.47 mmol l-1. Embryo development to the eight-cell stage was slightly enhanced 82% (14 of 17) versus 60% (24 of 40) when no glucose was added to the medium, and the resulting blastocysts had significantly more cells (99.1 +/- 13.5 versus 58.4 +/- 8.2; P < 0.02) than did embryos grown in the presence of 1 mmol glucose l-1.(ABSTRACT TRUNCATED AT 250 WORDS)
尽管人类胚胎能够在体外发育6天或更长时间,但对于主要营养物质丙酮酸和葡萄糖对胚胎发育的影响却知之甚少。由于胚胎在植入前发育过程中的营养需求会发生变化,因此研究人员使用从二细胞至四细胞阶段培养至囊胚阶段的“多余”人类植入前胚胎,在含有不同浓度丙酮酸和葡萄糖的培养基中,检测了改变培养基中底物浓度的影响。在单细胞阶段至二细胞至四细胞阶段,所有胚胎均暴露于0.47 mmol/L丙酮酸和5.5 mmol/L葡萄糖中。培养基中仅以丙酮酸作为底物时,囊胚发育率可达59%(17个中有10个)。相反,在无丙酮酸的培养基中培养胚胎,84%(25个中有21个)的胚胎发育停滞,对于16%(25个中有4个)发育至囊胚阶段的胚胎,在桑葚胚至囊胚阶段过渡的第4 - 5天,其代谢活性显著降低。胚胎无法利用葡萄糖来弥补培养基中丙酮酸的缺乏。丙酮酸摄取与外源浓度相关,在测试的最高浓度0.47 mmol/L时发生最佳发育。当培养基中不添加葡萄糖时,胚胎发育至八细胞阶段的比例略有提高,为82%(17个中有14个),而添加1 mmol/L葡萄糖时为60%(40个中有24个),并且所得囊胚的细胞数量显著更多(分别为99.1±13.5和58.4±8.2;P<0.02)。(摘要截取自250字)