CONICET, Instituto de Química y Fisicoquímica Biológicas Prof. Alejandro C. Paladini (IQUIFIB), Universidad de Buenos Aires, Buenos Aires, Argentina.
The Miami Project to Cure Paralysis and the Department of Neurological Surgery, University of Miami Miller School of Medicine, Miami, FL, USA.
Transgenic Res. 2018 Apr;27(2):135-153. doi: 10.1007/s11248-018-0062-5. Epub 2018 Feb 16.
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, S100β and p75) 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 S100β. 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.
骨髓单核细胞（BMMC）构成具有促进组织再生潜力的异质群体。出于这个原因，这种细胞群最近已成为间充质干细胞的治疗替代物，因为不需要培养并且可以避免表型转化。在这项工作中，为了实现长期的细胞标记和研究更长的存活时间，我们使用表达 GFP 的成年转基因大鼠分离的 BMMC 来复制我们的野生型模型，并在 Wallerian 变性的可逆模型中评估它们的髓鞘再生能力。RT-PCR 和流式细胞术分析证实，与野生型 BMMC 相比，从转基因株系分离的细胞表现出类似的多能祖细胞（CD34、CD90 和 CD105）和施万细胞（MPZ、MBP、S100β 和 p75）特异性标记物的表达水平。表达 GFP 的 BMMC 保持其迁移能力，仅到达受损的神经。最重要的是，通过长期的细胞追踪检测到，其中一些 BMMC 定居在脱髓鞘区域，与内源性细胞混合，经历表型变化并与施万细胞标记物 MBP 和 S100β 共定位。同样值得强调的是，转基因 BMMC 在 MBP 组织和水平方面复制了野生型 BMMC 的作用。基于这些发现，从转基因动物分离的 BMMC 构成了评估它们在外周神经系统脱髓鞘-髓鞘再生及其潜在机制中的作用的有用工具。
