| 大鼠心肌条件培养基对ICR小鼠胚胎干细胞分离克隆的影响 |
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| 引用本文： | 薛亚梅,王锋.大鼠心肌条件培养基对ICR小鼠胚胎干细胞分离克隆的影响[J].畜牧兽医学报,2005,36(5):440-445. |
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| 作者姓名： | 薛亚梅 王锋 |
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| 作者单位： | 南京农业大学动物科技学院动物胚胎工程技术中心,南京,210095 |
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| 基金项目： | 江苏省自然科学基金(BK2003075) |
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| 摘 要： | 采用大鼠心肌条件培养基(RH CM)培养ICR小鼠的桑椹胚和囊胚,发现由囊胚分离的ES细胞传代后ES集落的出现率显著高于桑椹胚(P<0.05),囊胚更适合作为ES细胞分离克隆的材料。以RH CM为培养基的试验组ES细胞传代的平均时间间隔为38 h,对照组传代的时间间隔平均为78 h,两者差异显著(P<0.05)。表明RH CM能够促进ES细胞贴壁增殖和ES集落的形成,有效地维持ES细胞未分化状态。试验中设计的3 种培养条件对原代ES集落的形成影响不显著,但对传代后的ES集落的形成和传代的代次有显著差异。其中以MEF作饲养层,添加RH CM培养基的效果最好。
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| 关 键 词： | 条件培养基 分离克隆 小鼠胚胎干细胞 心肌 大鼠 ES细胞 时间间隔 ICR小鼠 差异显著 形成影响 培养条件 桑椹胚 集落 囊胚 出现率 试验组 RH 饲养层 MEF 平均 CM 增殖 代次 添加 |
| 文章编号： | 0366-6964(2005)05-0440-06 |
The Effect of Rat Heart Conditioned Medium in Isolation and Cloning of Embryonic Stem Cells from ICR Mouse Embryo |
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| XUE Ya-mei,WANG Feng.The Effect of Rat Heart Conditioned Medium in Isolation and Cloning of Embryonic Stem Cells from ICR Mouse Embryo[J].Acta Veterinaria et Zootechnica Sinica,2005,36(5):440-445. |
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| Authors: | XUE Ya-mei WANG Feng |
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| Abstract: | ICR mouse blastocysts and morulas were cultured in rat heart conditioned medium (RH-CM).As far as the following indexes attaching ratio,ICM growth ratio and ES clone formation ratio are concerned,the choice of blastocysts is much better than morulas (P<0.05),therefore mouse blastocysts were more suitable than morulas to get ES cells. Interval between two passages (38 h) of expriment groups was significant higher than control group (P<0.05). It was showed that RH-CM not only could maintain ES cells in the undifferentiated state, inhibiting the spontaneous differentiation effectively, but also promote adherence and proliferation of mouse ES cells. The effect of the three culture systems on formation of ES colonies at primary passage was not remarkable but significant in ES colonies formation after passage and the highest times of passage. The combination of RH-CM with mitotically inactive MEF feeder layer was the best culture system. |
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| Keywords: | rat heart conditioned medium morula blastocyst ES cells |
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