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EGFP transgene: a useful tool to track transplanted bone marrow mononuclear cell contribution to peripheral remyelination

机译:EGFP转基因:一种用于跟踪移植骨髓单核细胞对外围重新髓鞘的有用工具

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Bone marrow mononuclear cells (BMMC) constitute a heterogeneous population with potential to promote tissue regeneration. For this reason, this cell fraction has recently become a therapeutic alternative to mesenchymal stem cells, as culture is not required and phenotypic transformations can be hence avoided. In this work, and in order to attain long-lasting cell labeling and study longer survival times, we used BMMC isolated from adult transgenic rats expressing GFP to reproduce our wild type model and evaluate their remyelination ability in a reversible model of Wallerian degeneration. RT-PCR and flow cytometry analysis confirmed that cells isolated from the transgenic strain exhibited similar expression levels of markers specific to multipotent progenitors (CD34, CD90 and CD105) and Schwann cells (MPZ, MBP, S100 beta and p75(NTR)) compared to wild type BMMC. BMMC expressing GFP retained their migration capacity, arriving exclusively at the injured nerve. Most importantly, and as detected through long-lasting cell tracking, some of these BMMC settled in the demyelinated area, mingled with endogenous cells, underwent phenotypic changes and colocalized with Schwann cell markers MBP and S100 beta. Also worth highlighting, transgenic BMMC replicated wild type BMMC effects in terms of MBP organization and levels. On the basis of these findings, BMMC isolated from transgenic animals constitute a useful tool to evaluate their role in peripheral nervous system demyelination-remyelination and the underlying mechanisms.
机译:骨髓单核细胞(BMMC)构成具有促进组织再生的潜力的异质群。因此,该细胞级分最近成为间充质干细胞的治疗替代物,因为不需要培养,因此可以避免表型转化。在这项工作中,为了获得持久的细胞标记和研究更长的存活时间,我们使用BMMC与表达GFP的成年转基因大鼠分离,以再现我们的野生型模型,并评估Wallerian变性可逆模型中的重新髓鞘化能力。 RT-PCR和流式细胞术分析证实,与基于野生型BMMC。表达GFP的BMMC保留了其迁移能力,专门在受伤的神经中抵达。最重要的是,并且通过长持久的单元跟踪检测,这些BMMC中的一些沉降在脱髓鞘区域中,与内源细胞混合,接受表型变化并与Schwann细胞标记物MBP和S100β结合。还值得突出显示,转基因BMMC在MBP组织和水平方面复制野生型BMMC效应。在这些发现的基础上,从转基因动物中分离的BMMC构成了一种有用的工具,以评估它们在外周神经系统脱髓鞘髓鞘中的作用和潜在机制。

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