Abstract: We reported here a new method of culturing and isolating ES cell colonies from mouse blastocysts. Monolayer culture was made by the heart tissue of a 2-3 weeks postnatal rat, and the conditioned medium (RHCM) were collected from cultures before passage 6. C57BL/6J mouse blastocysts were cultured in RHCM (without feeder layer), while the control group used primary mouse embryo fibroblast (ME) as feeder layer, with LIF added into the medium. After the blastocysts being cultured in RHCM, the rates of embryo attaching, ICM propagating and ES cell colonies forming, were 67%, 93% and 13% respectively. They had no prominent differences with those of the control group. This new method could be useful in establishing mouse ES cell lines simply and effectively.

Key words: rat heart, conditioned medium, mouse embryo, ES cell

摘要： 报道一种新的从小鼠囊胚中培养和分离出ES细胞集落的方法。取出生2～3周龄大鼠的心肌组织制备单层培养物，收集6代以内的条件培养基(RHCM)。以RHCM培养C57BL/6J小鼠的囊胚(无饲养层)，对照组以小鼠胚胎原代成纤维细胞(ME)为饲养层，并补加LIF因子。经过190个胚的实验证明，以RHCM为培养基培养小鼠囊胚时，胚胎贴壁率、ICM增殖率和ES细胞集落的出现率与对照组没有显著性差异，分别为67%、93%和13%。这种新的方法为简便而有效地建立与培养小鼠ES细胞系提供了有益的资料。

关键词: 大鼠心肌, 条件培养基, 小鼠胚胎, ES细胞