葡萄糖对ICR小鼠胚胎体外发育的影响  

Effects of Glucose on Development of ICR Mouse Embryos in vitro

作　　者：丁芳[1] 周红林[2] 刘洋[2] 马兰[2] 苏莹[2] 杜玲[2] 

机构地区：[1]三峡大学仁和医院妇产科,湖北宜昌443001 [2]昆明医学院第二附属医院妇科,云南昆明650101

出　　处：《Zoological Research》2007年第5期501-506,共6页动物学研究（英文）

基　　金：留学人员科技活动项目择优资助项目

摘　　要：研究葡萄糖在小鼠早期胚胎体外发育中的作用。实验1将6—8周龄的ICR雌鼠超数排卵后与公鼠交配,收集1-细胞放入含0(对照组)、0.5、1、3、5、10mmol/L葡萄糖的CZB中培养;实验2将从超排的ICR雌鼠输卵管内收集的1-细胞放入无糖CZB中培养,分别于1细胞、2细胞、4细胞、桑椹胚阶段移入含3.0mmol/L葡萄糖(最适浓度)的CZB中,培养24h后又移回到无糖CZB中(桑椹胚阶段除外)继续培养以及整个胚胎培养过程均在含糖CZB中,对照组胚胎培养全程均在无糖CZB中。每组胚胎于37℃、5%CO2培养箱中培养120h,每24h在倒置显微镜下观察胚胎发育情况,分别计算2-细胞率、4-细胞率、桑椹胚率、囊胚率和孵化率,并进行囊胚细胞计数。结果显示,小鼠胚胎在含糖CZB中与在无糖CZB中4-细胞发育率无差异;含糖CZB中囊胚率显著高于对照组;3.0mmol/L浓度组囊胚细胞数显著高于其余组;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高于对照组,1-细胞至2-细胞、桑椹胚及其以后阶段添加葡萄糖囊胚率与对照组无差异。实验证实,在ICR小鼠胚胎体外培养中加入葡萄糖不会导致2-细胞阻滞;葡萄糖浓度增至10mmol/L对ICR小鼠胚胎无毒性作用;ICR小鼠胚胎体外培养的最适葡萄糖浓度为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。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 10mmol/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 3.0mmol/L glucose （optimal concentration） and removed to glucose-free CZB 24h later （except the morula stage embryo）. The embryos in another group were cultured in CZB supplemented with 3.0mmol/L glucose during the entire culture time. The embryos in the control group were continuously cultured in glucose-free CZB. Embryos were cultured for 120h at 37 ℃ under 5% CO2 in sealed culture chambers and observed every 24h under a Nikon inverted microscope. The culture efficiency was evaluated by determining the proportion of embryos reaching the four-cell （48h）, blastocyst （96h）, or hatched blastocyst （120h） 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 3.0mmol/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

关 键 词：葡萄糖 小鼠 2-细胞阻滞 胚胎 体外发育 

分 类 号：Q813[生物学—生物工程]