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。
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.
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2005年第5期440-445,共6页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
江苏省自然科学基金(BK2003075)
