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Monitoring stem cell differentiation using Raman microspectroscopy: chondrogenic differentiation, towards cartilage formation
Analyst ( IF 3.6 ) Pub Date : 2020-11-02 , DOI: 10.1039/d0an01983f Francesca Ravera 1, 2, 3, 4, 5 , Esen Efeoglu 4, 6, 7, 8 , Hugh J. Byrne 2, 3, 4, 5
Analyst ( IF 3.6 ) Pub Date : 2020-11-02 , DOI: 10.1039/d0an01983f Francesca Ravera 1, 2, 3, 4, 5 , Esen Efeoglu 4, 6, 7, 8 , Hugh J. Byrne 2, 3, 4, 5
Affiliation
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Mesenchymal Stem Cells (MSCs) have the ability to differentiate into chondrocytes, the only cellular components of cartilage and are therefore ideal candidates for cartilage and tissue repair technologies. Chondrocytes are surrounded by cartilage-like extracellular matrix (ECM), a complex network rich in glycosaminoglycans, proteoglycans, and collagen, which, together with a multitude of intracellular signalling molecules, trigger the chondrogenesis and allow the chondroprogenitor to acquire the spherical morphology of the chondrocytes. However, although the mechanisms of the differentiation of MSCs have been extensively explored, it has been difficult to provide a holistic picture of the process, in situ. Raman Micro Spectroscopy (RMS) has been demonstrated to be a powerful analytical tool, which provides detailed label free biochemical fingerprint information in a non-invasive way, for analysis of cells, tissues and body fluids. In this work, RMS is explored to monitor the process of Mesenchymal Stem Cell (MSC) differentiation into chondrocytes in vitro, providing a holistic molecular picture of cellular events governing the differentiation. Spectral signatures of the subcellular compartments, nucleolus, nucleus and cytoplasm were initially probed and characteristic molecular changes between differentiated and undifferentiated were identified. Moreover, high density cell micromasses were cultured over a period of three weeks, and a systematic monitoring of cellular molecular components and the progress of the ECM formation, associated with the chondrogenic differentiation, was performed. This study shows the potential applicability of RMS as a powerful tool to monitor and better understand the differentiation pathways and process.
中文翻译：
使用拉曼光谱法监测干细胞分化：软骨分化，向软骨形成
间充质干细胞（MSC）具有分化为软骨细胞的能力，软骨细胞是软骨的唯一细胞成分，因此是软骨和组织修复技术的理想候选者。软骨细胞被软骨样细胞外基质（ECM）包围，该基质富含糖胺聚糖，蛋白聚糖和胶原蛋白，并与大量的细胞内信号分子一起触发软骨形成，并使软骨生成剂获得软骨的球形形态。软骨细胞。然而，尽管已经广泛探索了MSC分化的机制，但是难以原位提供该过程的整体图景。拉曼显微光谱（RMS）已被证明是功能强大的分析工具，它以无创方式提供详细的无标签生化指纹信息，用于分析细胞，组织和体液。在这项工作中，探索了RMS以监测间充质干细胞（MSC）体外分化为软骨细胞的过程，提供控制分化的细胞事件的整体分子图。最初探测亚细胞区室，核仁，细胞核和细胞质的光谱特征，并鉴定分化和未分化之间的特征性分子变化。此外，在三周的时间内培养了高密度的细胞微团，并进行了系统的细胞分子成分和与软骨形成分化相关的ECM形成过程的监测。这项研究表明RMS作为监视和更好地理解分化途径和过程的有力工具的潜在适用性。
更新日期：2020-11-06
中文翻译：
使用拉曼光谱法监测干细胞分化：软骨分化，向软骨形成
间充质干细胞（MSC）具有分化为软骨细胞的能力，软骨细胞是软骨的唯一细胞成分，因此是软骨和组织修复技术的理想候选者。软骨细胞被软骨样细胞外基质（ECM）包围，该基质富含糖胺聚糖，蛋白聚糖和胶原蛋白，并与大量的细胞内信号分子一起触发软骨形成，并使软骨生成剂获得软骨的球形形态。软骨细胞。然而，尽管已经广泛探索了MSC分化的机制，但是难以原位提供该过程的整体图景。拉曼显微光谱（RMS）已被证明是功能强大的分析工具，它以无创方式提供详细的无标签生化指纹信息，用于分析细胞，组织和体液。在这项工作中，探索了RMS以监测间充质干细胞（MSC）体外分化为软骨细胞的过程，提供控制分化的细胞事件的整体分子图。最初探测亚细胞区室，核仁，细胞核和细胞质的光谱特征，并鉴定分化和未分化之间的特征性分子变化。此外，在三周的时间内培养了高密度的细胞微团，并进行了系统的细胞分子成分和与软骨形成分化相关的ECM形成过程的监测。这项研究表明RMS作为监视和更好地理解分化途径和过程的有力工具的潜在适用性。
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