Sadan Ofer, Shemesh Noam, Barzilay Ran, Bahat-Stromza Merav, Melamed Eldad, Cohen Yoram, Offen Daniel
Department of Neurology, Sackler Faculty of Medicine, Laboratory of Neurosciences, Felsenstein Medical Research Center, Petah Tikva, Israel.
Stem Cells. 2008 Oct;26(10):2542-51. doi: 10.1634/stemcells.2008-0240. Epub 2008 Jul 17.
Stem cell-based treatment is a promising frontier for neurodegenerative diseases. We propose a novel protocol for inducing the differentiation of rat mesenchymal stem cells (MSCs) toward neurotrophic factor (NTF)-secreting cells as a possible neuroprotective agent. One of the major caveats of stem cell transplantation is their fate post-transplantation. To test the viability of the cells, we tracked the transplanted cells in vivo by magnetic resonance imaging (MRI) scans and validated the results by histology. MSCs went through a two-step medium-based differentiation protocol, followed by in vitro characterization using immunocytochemistry and immunoblotting analysis of the cell media. We examined the migratory properties of the cells in the quinolinic acid (QA)-induced striatal lesion model for Huntington's disease. The induced cells were labeled and transplanted posterior to the lesion. Rats underwent serial MRI scans to detect cell migration in vivo. On the 19th day, animals were sacrificed, and their brains were removed for immunostaining. Rat MSCs postinduction exhibited both neuronal and astrocyte markers, as well as production and secretion of NTFs. High-resolution two-dimensional and three-dimensional magnetic resonance images revealed that the cells migrated along a distinct route toward the lesion. The in vivo MRI results were validated by the histological study, which demonstrated that phagocytosis had only partially occurred and that MRI could correctly depict the status of the migrating cells. The results show that these cells migrated toward a QA lesion and therefore survived for 19 days post-transplantation. This gives hope for future research harnessing these cells for treating neurodegenerative diseases. Disclosure of potential conflicts of interest is found at the end of this article.
基于干细胞的治疗是神经退行性疾病一个很有前景的前沿领域。我们提出了一种新方案，用于诱导大鼠间充质干细胞（MSCs）向分泌神经营养因子（NTF）的细胞分化，作为一种可能的神经保护剂。干细胞移植的一个主要问题是移植后它们的命运。为了测试细胞的活力，我们通过磁共振成像（MRI）扫描在体内追踪移植的细胞，并通过组织学验证结果。MSCs经历了一个基于培养基的两步分化方案，随后使用免疫细胞化学和细胞培养基的免疫印迹分析进行体外表征。我们在喹啉酸（QA）诱导的亨廷顿舞蹈病纹状体损伤模型中检测了细胞的迁移特性。诱导的细胞被标记并移植到损伤部位后方。大鼠接受系列MRI扫描以在体内检测细胞迁移。在第19天，处死动物并取出其大脑进行免疫染色。诱导后的大鼠MSCs表现出神经元和星形胶质细胞标记物，以及NTF的产生和分泌。高分辨率二维和三维磁共振图像显示细胞沿着一条独特的路径向损伤部位迁移。体内MRI结果通过组织学研究得到验证，该研究表明吞噬作用仅部分发生，并且MRI能够正确描绘迁移细胞的状态。结果表明这些细胞向QA损伤部位迁移，因此在移植后存活了19天。这为未来利用这些细胞治疗神经退行性疾病的研究带来了希望。潜在利益冲突的披露见本文末尾。
