非接触状态下小鼠胚胎干细胞在小鼠肝癌细胞H22诱导下分化的动态观察

目的 观察小鼠源性肿瘤细胞分泌的细胞因子对小鼠胚胎干细胞(ES cells)的诱导分化.方法 将 ES细胞形成的拟胚体分为诱导组和对照组.诱导组在加入小鼠肝癌细胞H22上清液制成的条件培养基中生长,对照组在去除白血病抑制因子(LIF)的ES细胞培养液中生长.于诱导分化的第7、9、12、15天在倒置显微镜下动态观察两组分化情况,并分别测量上清液中AFP的含量.结果 诱导分化的第7天观察到诱导组拟胚体细胞群落的中心和周边有较为典型的小鼠肝癌样细胞形成,第9、12、15天肝癌样细胞数量逐渐增加.对照组拟胚体向三胚层细胞分化,未见肝癌细胞形成.第9,12,15天诱导组和对照组AFP含量经方差分析,差异有统计学意义(P<0.05).结论 小鼠胚胎干细胞在小鼠肝癌细胞H22分泌的细胞因子的诱导下可以分化为肝癌细胞.

The inducement of mouse embryonic stem cells by the mouse cancer cell line H22 in non-contact state

Objective To observe the differentiation of mouse embryonic stem ceils (ESCs) induced by mouse cancer cytokines in vitro. Methods 3.5-days embryos were extracted, and the embryoid bodies (EBs) were divided into inducement and control groups. The ESCs in inducement group lived in DMEM supplemented with the supematant of mouse liver cancer H22, and those in control group lived in DMEM without leukemia inhibitor factor (LIF). Morphology index was used to observe the differentiation of ESCs and AFP antibody was used to detect the expression of hepatic protein such as AFP in the supernatant at 7th,gth, 12th, 15th day after the ESCs were induced. Results The typical mouse cancer cells could be seen in the center and outer of the EBs at 7th day in inducement group, and at 9th, 12th and 15th day,the number was increased gradually. There was statistically significant difference in AFP contents between inducement and control groups on the day 9,12 and 15 ( P < 0.05). Conclusion The mouse ESCs can be induced to liver cancer cells by the mouse cancer cytokines.