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人牙髓干细胞通过Olig2转录因子的表达分化为少突胶质前体细胞。

Human Dental Pulp Stem Cells Differentiate into Oligodendrocyte Progenitors Using the Expression of Olig2 Transcription Factor.

作者信息

Askari Nahid, Yaghoobi Mohammad Mehdi, Shamsara Mehdi, Esmaeili-Mahani Saeed

机构信息

National Institute of Genetic Engineering and Biotechnology, Tehran, Iran.

出版信息

Cells Tissues Organs. 2014;200(2):93-103. doi: 10.1159/000381668. Epub 2015 May 7.

Abstract

The helix-loop-helix transcription factor Olig2 is essential for lineage determination of oligodendrocytes. Differentiation of stem cells into oligodendrocytes and transplanting them is a novel strategy for the repair of different demyelination diseases. Dental pulp stem cells (DPSCs) are of great interest in regenerative medicine due to their potential for repairing damaged tissues. In this study, DPSCs were isolated from human third molars and transfected with the human Olig2 gene as a differentiation inducer for the oligodendrogenic pathway. Following the differentiation procedure, the expression of Sox2, NG2, PDGFRα, Nestin, MBP, Olig2, Oct4, glial fibrillary acidic protein and A2B5 as stage-specific markers was studied by real-time RT-qPCR, immunocytochemistry and Western blot analysis. The cells were transplanted into a mouse model of local sciatic damage by lysolecithin as a model for demyelination. Oligodendrocyte progenitor cells (OPCs) actively remyelinated and recovered the lysolecithin-induced damages in the sciatic nerve as revealed by treadmill exercise, the von Frey filament test and hind paw withdrawal in response to a thermal stimulus. Recovery of behavioral reflexes occurred 2-6 weeks after OPC transplantation. The results demonstrate that the expression of Olig2 in DPSCs reduces the expression of stem cell markers and induces the development of oligodendrocyte progenitors as revealed by the emergence of oligodendrocyte markers. DPSCs could be programmed into oligodendrocyte progenitors and considered as a simple and valuable source for the cell therapy of neurodegenerative diseases.

摘要

螺旋-环-螺旋转录因子Olig2对于少突胶质细胞的谱系确定至关重要。将干细胞分化为少突胶质细胞并进行移植是修复不同脱髓鞘疾病的一种新策略。牙髓干细胞(DPSCs)因其具有修复受损组织的潜力而在再生医学中备受关注。在本研究中,从人类第三磨牙中分离出DPSCs,并用人Olig2基因进行转染,作为少突胶质细胞生成途径的分化诱导剂。在分化过程之后,通过实时RT-qPCR、免疫细胞化学和蛋白质印迹分析研究了Sox2、NG2、PDGFRα、Nestin、MBP、Olig2、Oct4、胶质纤维酸性蛋白和A2B5作为阶段特异性标志物的表达。将这些细胞通过溶血卵磷脂移植到局部坐骨神经损伤的小鼠模型中,作为脱髓鞘模型。如通过跑步机运动、von Frey细丝试验以及对热刺激的后爪退缩所显示的,少突胶质细胞前体细胞(OPCs)积极地进行髓鞘再生并修复了坐骨神经中溶血卵磷脂诱导的损伤。OPC移植后2至6周出现行为反射的恢复。结果表明,DPSCs中Olig2的表达降低了干细胞标志物的表达,并诱导了少突胶质细胞前体细胞的发育,这通过少突胶质细胞标志物的出现得以揭示。DPSCs可以被编程为少突胶质细胞前体细胞,并被视为神经退行性疾病细胞治疗的一种简单且有价值的细胞来源。

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