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丁芳, 周红林, . 2007: 葡萄糖对小鼠胚胎体外发育的影响. 动物学研究, 28(5): 501-506.
引用本文: 丁芳, 周红林, . 2007: 葡萄糖对小鼠胚胎体外发育的影响. 动物学研究, 28(5): 501-506.
DING Fang, ZHOU Hong-lin, LIU Yang, MA Lan, SU Ying, DU Ling. 2007. Effects of Glucose on Development of ICR Mouse Embryosin vitro. Zoological Research, 28(5): 501-506.
Citation: DING Fang, ZHOU Hong-lin, LIU Yang, MA Lan, SU Ying, DU Ling. 2007. Effects of Glucose on Development of ICR Mouse Embryosin vitro. Zoological Research, 28(5): 501-506.

葡萄糖对小鼠胚胎体外发育的影响

Effects of Glucose on Development of ICR Mouse Embryosin vitro

  • 摘要: 通过在CZB培养液中添加不同浓度葡萄糖及在胚胎发育的不同阶段加入葡萄糖,对小鼠胚胎进行体外培养,以探讨葡萄糖在小鼠早期胚胎体外发育中的作用。其结果表明,小鼠胚胎在含糖CZB与在无糖CZB中培养比较,4-细胞发育率无差异;各浓度葡萄糖组囊胚率显著高于无糖组,其中3.0mmol/L浓度组囊胚细胞数显著高于其余组;实验二:2-细胞至4-细胞、4-细胞至桑椹胚前添加葡萄糖囊胚率显著提高。上述结果证明,在小鼠胚胎体外培养中加入葡萄糖不会导致2-细胞阻滞;葡萄糖浓度增加至10mmol/L对小鼠胚胎无毒性作用,其最适浓度为3.0mmol/L;2-细胞至4-细胞、4-细胞至桑椹胚前添加葡萄糖是必要的。关键词 葡萄糖;小鼠;2-细胞阻滞;胚胎;体外发育

     

    Abstract: Abstract: To study the effects of glucose on the development of ICR mouse embryos in vitro. Experiment 1: ICR female mice (6-8 weeks of age) were super-ovulated with i.p. injections of PMSG and hCG, and mated overnight with males. One-cell embryos were collected at 22-26 hrs after hCG and cultured in CZB supplemented with 0, 0.5, 1.0, 3.0, 5.0 or 10 mmol/L glucose respectively. Experiment 2: One-cell embryos were collected from the oviducts of ICR female mice, super-ovulated and cultured in glucose-free CZB. The embryos,respectively at one-cell, two-cell, four-cell or morula stage,were removed from glucose-free CZB medium and placed in CZB medium supplemented with 3.0 mmol/L glucose (optimal concentration) and removed to glucose-free CZB 24 h later (except the morula stage embryo). The embryos in another group were cultured in CZB supplemented with 3.0 mmol/L glucose during the entire culture time. The embryos in the control group were continuously cultured in glucose-free CZB. Embryos were cultured for 120 h at 37℃ under 5% CO2 in sealed culture chambers and observed every 24 h under a Nikon inverted microscope. The culture efficiency was evaluated by determining the proportion of embryos reaching the four-cell (48 h), blastocyst (96 h), or hatched blastocyst (120 h) stage. The total cell numbers were counted in the blastocyst and hatched blastocyst embryos. The experiment results were: 1) There were no significant differences in the rates of four-cell embryos between every glucose-containing group and the glucose-free group. 2)The blastocyst rates of glucose-containing groups were significantly higher than that of the control. 3)The total cell numbers in the concentration group containing 3.0 mmol/L glucose were significantly higher than one another. 4)Addition of glucose from the two-cell to four-cell stage or from the four-cell to morula stage, significantly increased the blastocyst rates compared with the control. In contrast, addition of glucose from one-cell to two-cell, at morula or after morula, did not increase the blastocyst rates compared with the control. It indicates that addition of glucose to CZB does not result in two-cell block of the embryo development, and addition of glucose to CZB in a concentration as high as 10 mmol/L does not inhibit the development of ICR mouse one-cell to hatched blastocyst development. The optimal concentration of glucose added in CZB could be 3.0 mmol/L in the culture of ICR mouse embryos in vitro. Exposure of embryos to glucose, beginning at the two-cell and extending to the four-cell stage or beginning at the four-cell and extending to the morula stage, is necessary for the development of ICR mouse embryos in vitro.

     

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