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利用 SPR 生物传感器检测人骨髓间充质干细胞内皮分化过程中 VE-钙黏蛋白的早期表达。

Early-stage detection of VE-cadherin during endothelial differentiation of human mesenchymal stem cells using SPR biosensor.

机构信息

Research Center for Pharmaceutical Nanotechnology, Tabriz University of Medical Sciences, 51664-14766 Tabriz, Iran; Stem Cell Research Center, Tabriz University of Medical Sciences, 51664-14766 Tabriz, Iran.

Department of Applied Cell Sciences, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, 51664-14766 Tabriz, Iran.

出版信息

Biosens Bioelectron. 2017 Oct 15;96:358-366. doi: 10.1016/j.bios.2017.05.018. Epub 2017 May 10.

DOI:10.1016/j.bios.2017.05.018
PMID:28527412
Abstract

Surface plasmon resonance (SPR) biosensors are most commonly applied for real-time dynamic analysis and measurement of interactions in bio-molecular studies and cell-surface analysis without the need for labeling processes. Up to present, SPR application in stem cell biology and biomedical sciences was underused. Herein, a very simple and sensitive method was developed to evaluate human mesenchymal stem cells trans-differentiation to endothelial lineage of over a period of 14 days based on VE-cadherin biomarker. The SPR signals increased with the increase of the amount of VE-cadherin expression on the cell surface during cell differentiation process. The method was able to detect ≈27 cells permm. No significant effect was observed on the cell viability during the cell attachment to the surface of immune-reactive biochips and during the SPR analysis. Using this highly sensitive SPR method, it was possible to sense the early stage of endothelial differentiation on day 3 in label-free form, whereas flow cytometry and fluorescent microscopy methods were found unable to detect the cell differentiation at the same time. Therefore, the proposed method can rapidly and accurately detect cell differentiation in live cells and label-free manner without any need of cell breakage and has great potential for both diagnostic and experimental approaches.

摘要

表面等离子体共振(SPR)生物传感器最常用于实时动态分析和测量生物分子研究和细胞表面分析中的相互作用,而无需进行标记过程。到目前为止,SPR 在干细胞生物学和生物医学科学中的应用还未得到充分利用。本文开发了一种非常简单和灵敏的方法,基于 VE-钙黏蛋白生物标志物,在 14 天的时间内评估人骨髓间充质干细胞向血管内皮谱系的转分化。在细胞分化过程中,随着细胞表面 VE-钙黏蛋白表达量的增加,SPR 信号也随之增加。该方法能够检测到 ≈27 个细胞/平方毫米。在细胞附着到免疫反应性生物芯片表面和进行 SPR 分析的过程中,细胞活力没有受到显著影响。使用这种高灵敏度的 SPR 方法,可以在无标记的形式下在第 3 天检测到内皮分化的早期阶段,而流式细胞术和荧光显微镜方法在同一时间无法检测到细胞分化。因此,该方法可在无需细胞破裂的情况下,快速、准确地检测活细胞中的细胞分化,并具有很大的诊断和实验应用潜力。

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