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Abstract：
Embryonic stem cells (ESC) have the potential to generate homogeneous immature cells like stem/progenitor cells, which appear to be difficult to isolate and expand from primary tissue samples. In this study, we developed a simple method to generate homogeneous immature oligodendrocyte (OL) lineage cells from mouse ESC-derived neural stem cell (NSC). NSC converted to NG2+/OLIG2+double positive progenitors (NOP) after culturing in serum-free media for a week. NOP expressed Prox1, but not Gpr17 gene, highlighting their immature phenotype. Interestingly, FACS analysis revealed that NOP expressed proteins for NG2, but not PDGFRɑ, distinguishing them from primary OL progenitor cells (OPC). Nevertheless, NOP expressed various OL lineage marker genes including Cspg4, Pdgfrα, Olig1/2, and Sox9/10, but not Plp1 genes, and, when cultured in OL differentiation conditions, initiated transcription of Gpr17 and Plp1 genes, and expression of PDGFRα proteins, implying that NOP converted into a matured OPC phenotype. Unexpectedly, NOP remained multipotential, being able to differentiate into neurons as well as astrocytes under appropriate conditions. Moreover, NOP-derived OPC myelinated axons with a lower efficiency when compared with primary OPC. Taken together, these data demonstrate that NOP are an intermediate progenitor cell distinguishable from both NSC and primary OPC. Based on this profile, NOP may be useful for modeling mechanisms influencing the earliest stages of oligogenesis, and exploring the cellular and molecular responses of the earliest OL progenitors to conditions that impair myelination in the developing nervous system.
摘要：
胚胎干细胞（ESC）有可能产生同质的未成熟细胞，如干/祖细胞，似乎很难从原始组织样本中分离和扩增。在这项研究中,我们开发了一种简单的方法，从小鼠ESC衍生的神经干细胞（NSC）产生同质的未成熟少突胶质细胞（OL）谱系细胞。在无血清培养基中培养一周后,NSC转化为NG2+/OLIG2+双阳性祖细胞(NOP)。NOP表达Prox1，但不表达Gpr17基因，突出它们不成熟的表型。有趣的是,FACS分析显示NOP表达NG2蛋白，但不表达PDGFR,将它们与原代OL祖细胞(OPC)区分开。然而，NOP表达各种OL谱系标记基因，包括Cspg4，Pdgfrα，Olig1/2和Sox9/10，但不是Plp1基因，and,当在OL分化条件下培养时，Gpr17和Plp1基因的起始转录，和PDGFRα蛋白的表达，这意味着NOP转化为成熟的OPC表型。出乎意料的是,NOP保持多潜力，能够在适当的条件下分化成神经元和星形胶质细胞。此外，与原发性OPC相比，NOP衍生的OPC有髓轴突的效率较低。一起来看,这些数据证明NOP是可区别于NSC和原代OPC的中间祖细胞。根据这份资料,NOP可能对影响寡发生最早阶段的机制建模有用，并探索最早的OL祖细胞对损害发育中的神经系统髓鞘形成的条件的细胞和分子反应。
