ABSTRACT

Purpose: Previous studies have shown that oligodendrocytes and motor neurons have the same progenitors in the ventral spinal cord called spinal cord progenitor cells marked by oligodendrocyte lineage transcription factor 2 (Olig2). However, it is difficult to identify the spinal cord progenitor cell in vitro as they are present transiently and further transform into other neuronal (interneuron) and glial (oligodendrocyte) lineages during development. In the present study, we try to generated Olig2⁺ spinal cord progenitor cells from human induced neural stem cells (iNSCs) and identify those spinal cord progenitor cells in vitro

Materials and Methods: Human peripheral blood mononuclear cells (PBMCs) were converted into induced neural stem cells (iNSCs), after they were identified by immunostaining using neural stem cell markers such as Nestin, Sox1, Sox2, iNSCs were transformed into Olig2⁺ spinal cord progenitor cells in 3 weeks by using small molecules.

Results: Olig2⁺ spinal cord progenitor cells could expand for at least five passages and remained in a dividing state over a considerable period of time; in addition, the Olig2⁺ progenitor cells could mature into O4 and MBP positive oligodendrocytes and HB9 positive motor neurons in a short period.

Conclusion: Our research provides a useful protocol for rapid generation of human oligodendrocytes and motor neurons from human iNSCs and demonstrates a progenitor cell model for exploring the origin of motor neurons and oligodendrocyte in vitro, which will contribute to research on the development of spinal cord and regenerative medicine.

摘要

目的：以前的研究表明，少突胶质细胞和运动神经元在脊髓腹侧具有相同的祖细胞，称为脊髓祖细胞，由少突胶质细胞谱系转录因子 2 (Olig2) 标记。然而，很难在体外鉴定脊髓祖细胞，因为它们短暂存在并在发育过程中进一步转化为其他神经元（中间神经元）和神经胶质（少突胶质细胞）谱系。在本研究中，我们尝试从人诱导神经干细胞 (iNSCs) 中生成 Olig2 ⁺脊髓祖细胞，并在体外鉴定这些脊髓祖细胞

材料与方法：将人外周血单个核细胞（PBMCs）转化为诱导神经干细胞（iNSCs），通过免疫染色使用Nestin、Sox1、Sox2等神经干细胞标志物进行鉴定，将iNSCs转化为Olig2 ⁺脊髓祖细胞在 3 周内通过使用小分子。

结果： Olig2 ⁺脊髓祖细胞至少可以扩增5代，并在相当长的一段时间内保持分裂状态；此外，Olig2 ⁺祖细胞可在短时间内成熟为O4和MBP阳性少突胶质细胞和HB9阳性运动神经元。

结论：我们的研究为从人 iNSCs 中快速生成人少突胶质细胞和运动神经元提供了一个有用的方案，并展示了一种用于在体外探索运动神经元和少突胶质细胞起源的祖细胞模型，这将有助于研究脊髓和运动神经元的发育。再生医学。