Formation of embryoid bodies by mouse embryonic stem cells on plastic surfaces
Formation of embryoid bodies by mouse embryonic stem cells on plastic surfaces
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DOI：
10.1263/jbb.100.88
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发表时间：
2005-07-01
影响因子：
2.8
通讯作者：
Ito, Y
中科院分区：
文献类型：
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作者：
Konno, T;Akita, K;Ito, Y
将小鼠胚胎干（ES）细胞培养在具有磷脂聚合物（磷脂酰胆碱，PC）表面的人工高分子生物膜上。胚胎干细胞在播种后立即在PC表面聚集形成胚状体（EB）。3 d后在PC表面形成单个EBs，其大小取决于初始播种细胞的数量。相比之下，许多具有不均匀形状的小EBs在传统的疏水性未经处理的聚苯乙烯表面上形成。细胞的RT-PCR分析表明，与聚苯乙烯表面形成的EBs相比，在PC表面形成的EBs细胞间的相互作用增强。作为ES细胞向神经元分化标志的转录因子Pax6在PC表面形成的EBs中未表达；然而，在聚苯乙烯表面形成的EBs确实表达了Pax6，这表明它们正在向神经元分化。当受视黄酸（一种向神经元分化的诱导剂）刺激时，PC表面的EBs表达Pax6。我们还观察到胚胎干细胞与PC表面的粘附性降低。因此，大EBs在PC表面的形成是由于细胞间相互作用的增强和对神经元非特异性分化的抑制。
Mouse embryonic stem (ES) cells were cultured on artificial polymeric biomembranes with a phospholipid polymer (phosphatidylcholine, PC) surface. ES cells aggregated to form an embryoid body (EB) on the PC surface immediately after seeding. Single EBs formed on the PC surface after 3 d, and their size was depended on the initial number of cells that were seeded. In contrast, many small EBs with a nonuniform shape formed on a conventional hydrophobic nontreated polystyrene surface. RT-PCR assays of the Ells indicated that cell-cell interactions were enhanced in EBs that formed on the PC surface compared with EBs that formed on the polystyrene surface. The transcription factor Pax6, which is a marker of the differentiation of ES cells to neurons, was not expressed in EBs that formed on the PC surface; however, EBs that formed on the polystyrene surface did express Pax6, indicating that they were undergoing differentiation into neurons. When stimulated with retinoic acid (an inducer of differentiation into neurons), EBs on the PC surface expressed Pax6. We also observed that the adhesion of ES cells to the PC surface was reduced. Thus, the formation of large EBs on the PC surface was due to enhanced cell-cell interaction and inhibition of nonspecific differentiation to neurons.