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OALib Journal期刊
ISSN: 2333-9721
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体外诱导balb/c小鼠es细胞定向分化为胰岛样细胞团的形态观察
Keywords: 胚胎干细胞,定向分化,胰岛样细胞团,原子力显微镜,可视化
Abstract:
目的体外诱导balb/c小鼠胚胎干细胞定向分化为胰岛样细胞团,观察细胞表面形态学的变化。方法选用balb/c小鼠es细胞,经过拟胚体(eb)发育分化4d后开始定向诱导培养,不同时间段分别向细胞培养基中添加碱性成纤维细胞生长因子、胰岛素样生长因子-1和尼克酰胺和n等细胞生长因子,使es细胞定向分化为胰岛样细胞团。免疫细胞化学检测表达胰岛素和胰高血糖素的阳性细胞。原子力显微镜(afm)原位扫描阳性细胞团。结果eb细胞生长为大小不一、境界清楚的细胞团,细胞团内细胞排列紧密。胰岛b细胞数量多,主要分布在细胞团的中央,边缘少,染色较淡。而表达胰高血糖素的a细胞主要分布在细胞团的边缘,数量相对较少。afm扫描可见表达胰岛素的阳性细胞团,其表面有很多类似神经纤维的纤维束,连接成网络状。在细胞质中,还有很多类圆形的颗粒物质,其大小几乎一致,粒径都处于0.5~1.0μm间。结论诱导分化得到的细胞团不仅有形态和功能上的成熟,而且还具备良好的组织结构,为细胞的移植疗法提供了可靠的凭据。
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