AN IMPROVED CULTURE-MEDIUM SUPPORTS DEVELOPMENT OF RANDOM-BRED 1-CELL MOUSE EMBRYOS INVITRO
AN IMPROVED CULTURE-MEDIUM SUPPORTS DEVELOPMENT OF RANDOM-BRED 1-CELL MOUSE EMBRYOS INVITRO
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DOI：
10.1530/jrf.0.0860679
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发表时间：
1989-07-01
期刊：
影响因子：
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通讯作者：
TORRES, I
中科院分区：
文献类型：
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作者：
CHATOT, CL;ZIOMEK, CA;TORRES, I
单细胞CF-1 ×B6 SJLF 1/J胚胎，通常表现出2-细胞阻滞体外发育，已培养到囊胚期使用CZB培养基和葡萄糖洗涤程序。CZB培养基是改良的BMOC-2的进一步改良，含有116、1 mM-谷氨酰胺和0.1 mM-EDTA的增加的乳酸/丙酮酸比，但缺乏葡萄糖。在CZB培养基中连续培养单细胞胚胎，83%的胚胎发育超过2-细胞期，其中63%在培养72 h时为桑椹胚，但囊胚不发育。然而，在培养48小时（3-4细胞期）后将胚胎洗涤到含有葡萄糖的CZB培养基中足以允许发育进行，其中48%的胚胎在培养96小时时达到囊胚期。暴露于葡萄糖的胚胎是必要的，从3-4-细胞阶段通过早期桑椹胚阶段，因为冲洗回培养基CZB没有葡萄糖在72小时的培养仍然促进了50%的胚胎发育到囊胚期。在培养的前48小时（1-细胞至4-细胞阶段），该培养基中存在葡萄糖对胚胎发育不利。然而，谷氨酰胺对胚胎从1-细胞期发育到4-细胞期有有益的影响，尽管在培养的最后48小时内，它的存在不是发育进行的必要条件。在最佳条件下发育的囊胚平均含有33.7个总细胞。1-细胞胚胎的体外发育超过2-细胞阶段，以响应去除葡萄糖和添加谷氨酰胺到培养基中表明，葡萄糖可能会阻止一些基本的代谢过程，谷氨酰胺可能是这些小鼠胚胎早期发育过程中的首选能量底物。
One-cell CF-1 .times. B6SJLF1/J embryos, which usually exhibit a 2-cell block to development in vitro, have been cultured to the blastocyst stage using CZB medium and a glucose washing procedure. CZB medium is a further modification of modified BMOC-2 containing an increased lactate/pyruvate ratio of 116, 1 mM-glutamine and 0.1 mM-EDTA but lacking glucose. Continuous culture of one-cell embryos in CZB medium allowed 83% of embryos to develop beyond the 2-cell stage of which 63% were morulae at 72 h of cultured, but blastocysts did not develop. However, washing embryos into CZB medium containing glucose after 48 h of culture (3-4 cell stage) was sufficient to allow development to proceed, with 48% of embryos reaching the blastocyst stage by 96 h of culture. Exposure of embryos to glucose was only necessary from the 3-4-cell stage through the early morula stage since washing back into medium CZB without glucose at 72 h of culture still promoted the development of 50% of embryos to the blastocyst stage. The presence of glucose in this medium for the first 48 h of culture (1-cell to 4-cell stage) was detrimental to embryo development. Glutamine, however, exerted a beneficial effect on embryo development from the 1-cell to the 4-cell stage although its presence was not requiree for development to proceed during the final 48 h of culture. Blastocysts which developed under optimum conditions contained an average of 33.7 total cells. The in-vitro development of 1-cell embryos beyond the 2-cell stage in response to the removal of glucose and the addition of glutamine to the culture medium suggests that glucose may block some essential metabolic process, and that glutamine may be a preferred energy substrate during early development for these mouse embryos.