摘要
目的探讨小鼠胚胎体外发育中的最佳培养方案。方法将从超排的ICR雌鼠输卵管内收集的1-细胞放入无糖CZB中培养,分别于2-细胞、4-细胞、桑椹胚阶段更换入含3.0 mmol/L葡萄糖(最适浓度)的CZB中,以及胚胎培养全程均在含糖CZB中更换1次培养液、胚胎培养全程均在含糖CZB中不更换培养液,对照组胚胎培养全程均在无糖CZB中,观察记录胚胎的发育情况。结果2-细胞、4-细胞阶段换入含糖CZB中的序贯培养及培养全程在含糖CZB中单一培养,囊胚率均高于对照组(P<0.05);桑椹胚阶段换入含糖CZB中的序贯培养,囊胚率与对照组差异无统计学意义(P>0.05);2-细胞、4-细胞阶段换入含糖CZB中的序贯培养与全程在含糖CZB中的两步法单一培养及一步法单一培养,囊胚率分别为46.5%、38.4%、41.7%、56.6%,其差异无统计学意义(P>0.05)。结论在小鼠早期胚胎体外发育中,2-细胞至桑椹胚前对葡萄糖存在依赖性;尽管序贯培养是有效的,但并不一定优于一步单一培养。
Objective To seek the appropriate way for the culture of mouse early embryos in vitro. Methods One-cell embryos were collected from the oviducts of ICR female mice,super-ovulated and cultured in glucose-free CZB. The embryos, respectively at 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）. The embryos in the other tWO groups were cultured in CZB supplemented with 3.0 retool/ L glucose during the entire culture time with or without renewal of medium. The embryos in the control group were continuously cultured in glucose-free CZB. Embryo development was recorded a,t 24 h,48 h,72 h and 96 h respectively culture. Results （1）Shift embryos from glucose-free CZB to glucose-containing CZB at two-cell or four-cell stage, the blastocyst rates significantly increased compared with the control group. （2）The blastocyst rates of which the embryos were continuously cultured in glucose- containing CZB during the entire culture time with or without renewal of medium,were significantly higher than that of the control group. （3） Shift embryos from glucose-free CZB to glucose-containing CZB at morula stage, the hlastocyst rates did not increase compared with the control group. （4）The blastocysl rates,shift embryos from glucose-free CZB to glucose-containing CZB at two-cell or four-cell stage and continuously cultured in glucose-containing CZB during the entire culture time with or without renewal of medium,were 46.5%, 38.4% ,41.7% and 56.6% respectively, but there were no significant differences among groups. Conclusion Exposure of embryos to glucose, beginning at the two-cell and extending to the morula stage,is necessary for the development of ICR mouse embryos in vitro. Sequential Cuhure is sufficient,but is not necessarily superior to continuous Culture without renewal of medium.
出处
《重庆医学》
CAS
CSCD
北大核心
2010年第8期907-909,共3页
Chongqing medicine
关键词
小鼠
早期胚胎
培养方法
葡萄糖
序贯培养
单一培养
mouse
early embryo
cultural method
glucose
sequential culture
continuous culture
