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纳米颗粒标记大鼠骨髓间充质干细胞:其在分化和追踪中的应用

Nanoparticle labeling of bone marrow-derived rat mesenchymal stem cells: their use in differentiation and tracking.

作者信息

Akhan Ece, Tuncel Donus, Akcali Kamil C

机构信息

Department of Molecular Biology and Genetics, Bilkent University, 06800 Ankara, Turkey.

Department of Chemistry, Bilkent University, 06800 Ankara, Turkey.

出版信息

Biomed Res Int. 2015;2015:298430. doi: 10.1155/2015/298430. Epub 2015 Jan 14.

Abstract

Mesenchymal stem cells (MSCs) are promising candidates for cellular therapies due to their ability to migrate to damaged tissue without inducing immune reaction. Many techniques have been developed to trace MSCs and their differentiation efficacy; however, all of these methods have limitations. Conjugated polymer based water-dispersible nanoparticles (CPN) represent a new class of probes because they offer high brightness, improved photostability, high fluorescent quantum yield, and noncytotoxicity comparing to conventional dyes and quantum dots. We aimed to use this tool for tracing MSCs' fate in vitro and in vivo. MSC marker expression, survival, and differentiation capacity were assessed upon CPN treatment. Our results showed that after CPN labeling, MSC markers did not change and significant number of cells were found to be viable as revealed by MTT. Fluorescent signals were retained for 3 weeks after they were differentiated into osteocytes, adipocytes, and chondrocytes in vitro. We also showed that the labeled MSCs migrated to the site of injury and retained their labels in an in vivo liver regeneration model. The utilization of nanoparticle could be a promising tool for the tracking of MSCs in vivo and in vitro and therefore can be a useful tool to understand differentiation and homing mechanisms of MSCs.

摘要

间充质干细胞(MSCs)因其能够迁移至受损组织且不引发免疫反应,成为细胞治疗的理想候选者。目前已开发出多种技术用于追踪MSCs及其分化效果;然而,所有这些方法都存在局限性。基于共轭聚合物的水分散性纳米颗粒(CPN)代表了一类新型探针,因为与传统染料和量子点相比,它们具有高亮度、更高的光稳定性、高荧光量子产率以及无细胞毒性。我们旨在利用该工具在体外和体内追踪MSCs的命运。在CPN处理后评估了MSC标志物表达、存活率和分化能力。我们的结果表明,CPN标记后,MSC标志物未发生变化,MTT检测显示大量细胞存活。在体外分化为骨细胞、脂肪细胞和软骨细胞后,荧光信号保留了3周。我们还表明,在体内肝再生模型中,标记的MSCs迁移至损伤部位并保留了其标记。纳米颗粒的应用可能是体内外追踪MSCs的一种有前景的工具,因此可能是理解MSCs分化和归巢机制的有用工具。

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