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Mesenchymal stem cells cultured on magnetic nanowire substrates

机译：在磁性纳米线基材上培养的间充质干细胞

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Stem cells have been shown to respond to extracellular mechanical stimuli by regulating their fate through the activation of specific signaling pathways. In this work, an array of iron nanowires (NWs) aligned perpendicularly to the surface was fabricated by pulsed electrodepositon in porous alumina templates followed by a partial removal of the alumina to reveal 2-3 mu m of the NWs. This resulted in alumina substrates with densely arranged NWs of 33 nm in diameter separated by 100 nm. The substrates were characterized by scanning electron microscopy (SEM) energy dispersive x-ray analysis and vibrating sample magnetometer. The NW array was then used as a platform for the culture of human mesenchymal stem cells (hMSCs). The cells were stained for the cell nucleus and actin filaments, as well as immunostained for the focal adhesion protein vinculin, and then observed by fluorescence microscopy in order to characterize their spreading behavior. Calcein AM/ethidium homodimer-1 staining allowed the determination of cell viability. The interface between the cells and the NWs was studied using SEM. Results showed that hMSCs underwent a re-organization of actin filaments that translated into a change from an elongated to a spherical cell shape. Actin filaments and vinculin accumulated in bundles, suggesting the attachment and formation of focal adhesion points of the cells on the NWs. Though the overall number of cells attached on the NWs was lower compared to the control, the attached cells maintained a high viability (>90%) for up to 6 d. Analysis of the interface between the NWs and the cells confirmed the re-organization of F-actin and revealed the adhesion points of the cells on the NWs. Additionally, a net of filopodia surrounded each cell, suggesting the probing of the array to find additional adhesion points. The cells maintained their round shape for up to 6 d of culture. Overall, the NW array is a promising nanostructured platform for studying and influencing hMSCs
机译：干细胞已通过特定信号传导途径的激活调节他们的命运显示出对细胞外机械刺激响应。在这项工作中，垂直地取向的表面的铁纳米线（NWS）的阵列，通过脉冲电沉积在多孔氧化铝模板随后是部分除去氧化铝揭示2-3微米的纳米线的制造。这导致了具有直径为33nm密集地布置纳米线氧化铝基底相隔100纳米。通过扫描电子显微镜（SEM）的能量分散型X射线分析和振动样品磁衬底进行了表征。然后将NW阵列用作人类间充质干细胞（hMSCs）的培养物的平台。将细胞染色细胞核和肌动蛋白丝，以及免疫染色的粘着斑蛋白纽蛋白，然后以表征它们的铺展性能通过荧光显微镜观察。钙黄绿素AM /乙锭同型二聚体-1染色允许细胞活力的测定。细胞和纳米线之间的界面处使用SEM研究。结果表明，接受的hMSC的肌动蛋白丝的是翻译成从拉长至球形细胞形状的变化的重新组织。肌动蛋白丝和粘着斑蛋白在捆积累，这表明在纳米线的细胞粘着斑点的连接和形成。尽管附着在纳米线单元的整体数量相比要低的控制下，附着的细胞保持长达6 d高的存活率（> 90％）。该纳米线与细胞之间的接口进行的分析证实F-肌动蛋白的重新组织，并透露了关于纳米线细胞的粘附点。此外，丝状伪足的净包围的各小区，这表明该阵列的探测找到额外粘附点。将细胞保持其圆形形状长达6 d培养的。总体而言，NW阵列是研究和影响的hMSCs有前途的纳米结构平台

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