Brown J J, Whittingham D G
Medical Research Council, St George's Hospital Medical School, Tooting, London, UK.
Development. 1991 May;112(1):99-105. doi: 10.1242/dev.112.1.99.
Embryos of certain inbred mouse strains, and their F1 hybrids, are able to develop from the 1-cell to blastocyst stage in simple chemically defined media containing lactate (L), pyruvate (P) and glucose (G). The individual roles of these substrates in supporting complete preimplantation development in vitro was examined with 1-cell F2 embryos from B6CBF1 hybrid mice. Embryos collected between 26 and 27 h post hCG were cultured in medium containing L, P, LP or LPG. After 50 h in culture, the proportions developing to the morula stage were 1%, 83%, 94% and 100%, respectively. In combination, lactate and pyruvate appeared to act synergistically and both the rate and level of development to the morula stage were unaffected by the absence of glucose. After a further 46 h in culture, only the embryos grown in the presence of glucose developed into blastocysts. In LP medium, embryos arrested at the compacted morula stage late on day 3 of development. As culture continued in the absence of glucose, embryos decompacted (approximately 82 h post hCG) and subsequently degenerated. Exposure to medium containing glucose for the first, second or third 24 h period in culture was sufficient to support the morula-to-blastocyst transition. Glucose still supported this transition when embryos were transferred to LPG medium 3 h after the completion of compaction (76 h post hCG), but was ineffective 6 h later (82 h post hCG) once decompaction had commenced. We conclude that lactate and pyruvate together are able to support normal development of 1-cell F2 embryos to the morula stage in vitro, but that glucose is an essential component of the culture medium for development to the blastocyst stage.
某些近交系小鼠品系的胚胎及其F1杂种胚胎,能够在含有乳酸(L)、丙酮酸(P)和葡萄糖(G)的简单化学成分明确的培养基中从单细胞发育到囊胚阶段。利用B6CBF1杂种小鼠的单细胞F2胚胎,研究了这些底物在支持体外完全着床前发育中的各自作用。在hCG注射后26至27小时收集的胚胎,在含有L、P、LP或LPG的培养基中培养。培养50小时后,发育到桑椹胚阶段的比例分别为1%、83%、94%和100%。乳酸和丙酮酸联合作用时似乎具有协同效应,并且在没有葡萄糖的情况下,发育到桑椹胚阶段的速度和程度均不受影响。再培养46小时后,只有在有葡萄糖存在的情况下生长的胚胎发育成囊胚。在LP培养基中,胚胎在发育第3天后期停滞在致密桑椹胚阶段。随着在无葡萄糖条件下继续培养,胚胎开始松散(约在hCG注射后82小时),随后退化。在培养的第一个、第二个或第三个24小时期间接触含葡萄糖的培养基足以支持桑椹胚向囊胚的转变。当胚胎在致密化完成后3小时(hCG注射后76小时)转移到LPG培养基中时,葡萄糖仍能支持这种转变,但一旦开始松散(hCG注射后82小时)6小时后则无效。我们得出结论,乳酸和丙酮酸共同能够支持单细胞F2胚胎在体外正常发育到桑椹胚阶段,但葡萄糖是发育到囊胚阶段的培养基的必需成分。