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从家兔早期胚胎分离ES细胞影响因素的研究
Research on factors related to isolation of embryo stem cells from primary rabbit embryos

作  者: ; ; (王新庄); (王轶敏); (牛晖);

机构地区: 华南农业大学兽医学院

出  处: 《广西农业生物科学》 2006年第B09期68-72,共5页

摘  要: 比较胚胎的不同处理方法,不同饲养层对兔胚胎内细胞团(ICM)贴壁和增殖的影响。结果显示:消化掉外层粘蛋白和部分透明带的胚胎96 h贴壁率(85.29%,29/34)较高,内细胞团生长良好;超排组胚胎的贴壁率和内细胞团(ICM)原代克隆率与自然发情组差异不显著(85.71%vs 88.06%,21.43%vs19.40%,P>0.05);与对照组(12.5%,3.13%)相比,在小鼠胚胎成纤维细胞饲养层(M EF)和兔胚胎成纤维细胞饲养层(REF)上培养获得的呈聚集状生长ICM及ICM原代克隆率较高(P<0.01),M EF组和REF组差异不显著(54.1%,22.95%vs 51.92%,17.31%,P>0.05),但REF饲养层培养效果不稳定。研究结果表明,兔早期胚胎消化掉外层粘蛋白和部分透明带有利于胚胎内细胞团贴壁和增殖;超排处理不影响从兔胚中分离胚胎干细胞(ES);小鼠胚胎成纤维细胞饲养层能有效支持兔胚内细胞团的贴壁和增殖。 Different treatments of rabbit blastocyst and different feed layers were adopted to compare the effects on attachment and reproduction of rabbit inner cell mass (ICM). Results: There was high rate of attachment of blastocysts that were indigested external mucoprotein and partial zona pellucida in 96 h culture (85.29%, 29/34) and growth of ICM was excellent; there was not significant difference in rate of attachment and clone of primary ICM between superovulation group and contrast group (85.71% vs 88.06%, 21.43% vs 19.40%, P〉0.05); ratio of aggregated ICM and clone of primary ICM on mouse embryonic fibroblast (MEF) and rabbit embryonic fibroblast (REF) were higher compared with contrast group, but there was not significant difference (54.1%, 22.95% vs 51.92%, 17.31%, P〉0.05) in experimental groups (12.5%, 3.13% ). Conclusion:Rabbit blastocysts indigested external mucoprotein and partial zona pellucida are easy to attachment and reproduction;superovulation has no effect on in vitro isolation of rabbit embryo stem cells; it is effective to ICM attachment and reproduction on MEF.

关 键 词: 胚胎 胚胎干细胞 内细胞团 体外培养

领  域: [农业科学] [农业科学] [农业科学]

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