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不同体外培养条件对家兔早期囊胚贴壁行为的影响
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夏霞1，2 ,王新庄1,2,曹君君2，4,王轶敏2，3,牛晖2
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(1.河南农业大学 牧医工程学院，河南 郑州450002； 2.河南省肉牛工程技术研究中心,
河南 郑州450003； 3.西北农林科技大学 动物科技学院，陕西 杨凌712100；
4.华南农业大学 兽医学院，广东 广州510642）
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摘要：
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采集家兔自然交配后96h的早期囊胚，以低糖DMEM+150mL/L胎牛血清+0.1mmol/L非必需氨基酸+100IU/mL青霉素+100IU/mL链霉素为基础培养基，比较了不同胚胎处理方法、不同饲养层以及培养液中添加不同成分对兔早期囊胚贴壁和增殖的影响，以完善兔胚胎干细胞的建系方法。结果表明，以胚胎分割法和链霉蛋白酶-E(proteinase E)处理掉黏蛋白及部分透明带的胚胎容易贴壁和增殖；在小鼠成纤维细胞饲养层和兔胎儿成纤维细胞饲养层上，胚胎的脱带率差别不大，但在小鼠成纤维细胞饲养层上贴壁率明显提高，且贴壁后内细胞团增殖较快；添加胰岛素、白血病抑制因子和β-巯基乙醇均利于抑制ES的分化和促进内细胞团的增殖。 |
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关键词：
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胚胎干细胞；体外培养；胚胎分割；囊胚；家兔 |
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中图分类号：S 852.1 文献标识码：
A 文章编号：1673-4696(2006)07-0561-05 |
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Effect of different conditions of in vitro cultivation on attachment
of rabbit primary blastocyst |
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XIA Xia1,2, WANG Xin-zhuang1,2, CAO
Jun-jun2，4, WANG Yi-min2,3, NIU Hui2 |
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(1.College of Animal and Veterinary Medicine Engineering, Henan Agricultural University, Zhengzhou
450002,China;2.Research Center of Beef Engineering, Zhengzhou 450003, China;
3.College of Animal Science and Technology, Northwest University of Agriculture and Forestry, Yangling 712100,
China;4.College of Veterinary Medicine, South China Agricultural University,Guangzhou 510642,China) |
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Abstract: Early blastocysts were collected from rabbit at 96th h after mating. Effects of different treatments, feeders, media ingredients on attachment and reproduction of rabbit primary blastocysts were compared by using the basic media consisting of low glucose DMEM,15% of FBS ,0.1mmol/L of non-essential amino acids, 100IU/mL of penicillin and 100IU/mL of streptomycin. Embryo from division and treatment of proteinase E were good at attachment and reproduction. There was no significant difference between mouse fiberoblast cell feeder and rabbit embryonic fiberoblast cell feeder, but embryos cultured in mouse fiberoblast cell feeder had higher attachment and reproduction rates. The media supplied respectively with insulin, leukaemia inhibitory factor and β- mercaptoethanol were good at inhibiting the differentiation of ES-like cells and promoting reproduction of ICM. |
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Key words: embryonic stem cells; in vitro culture; embryonic partition; blastocysts; rabbit |
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