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Raman and infrared spectroscopy differentiate senescent from proliferating cells in a human dermal fibroblast 3D skin model
Analyst ( IF 4.2 ) Pub Date : 2017-08-07 00:00:00 , DOI: 10.1039/c7an00592j
Katharina Eberhardt 1, 2, 3, 4, 5 , Christian Matthäus 1, 2, 3, 4, 5 , Doreen Winter 3, 6, 7, 8 , Cornelia Wiegand 3, 6, 7, 8 , Uta-Christina Hipler 3, 6, 7, 8 , Stephan Diekmann 2, 3, 9, 10 , Jürgen Popp 1, 2, 3, 4, 5
Affiliation  

Senescent cells contribute to tissue aging and dysfunction. Therefore, detecting senescent cells in skin is of interest for skin tumor diagnostics and therapy. Here, we studied the transition into senescence of human dermal fibroblasts (HDFs) in a three-dimensional (3D) human fibroblast-derived matrix (FDM). Senescent and proliferating cells were imaged by Raman spectroscopy (RS) and Fourier transform infrared (FTIR) spectroscopy. The obtained averaged spectra were analyzed using PLS-LDA. For these 3D cultured cells, RS and FTIR could clearly distinguish senescent from proliferating cells. For both techniques, we detected senescence-associated alterations in almost all cellular macromolecules. Furthermore, we identified different biochemical properties of 3D compared to two-dimensional (2D) cultured cells, indicating that cells in their natural, skin-like 3D environment act differently than in (2D) cell cultivations in vitro. Compared to 2D cultured cells, cells grown in 3D models displayed a sharper contrast between the proliferating and senescent state, also affecting the abundance of biomolecules including nucleic acids. The training accuracies of both vibrational spectroscopic techniques were >96%, demonstrating the suitability of these label-free measurements for detecting these cellular states in 3D skin models.

中文翻译:

拉曼光谱和红外光谱在人类皮肤成纤维细胞3D皮肤模型中将衰老与增殖细胞区分开来

衰老细胞导致组织衰老和功能障碍。因此,检测皮肤中的衰老细胞对于皮肤肿瘤的诊断和治疗是令人感兴趣的。在这里,我们研究了在三维(3D)人类成纤维细胞衍生基质(FDM)中人类真皮成纤维细胞(HDF)衰老的过渡。通过拉曼光谱(RS)和傅立叶变换红外光谱(FTIR)光谱对衰老和增殖细胞成像。使用PLS-LDA分析获得的平均光谱。对于这些3D培养的细胞,RS和FTIR可以清楚地区分衰老与增殖细胞。对于这两种技术,我们检测了几乎所有细胞大分子中与衰老相关的改变。此外,与二维(2D)培养的细胞相比,我们确定了3D的不同生化特性,这表明细胞处于自然状态,体外。与2D培养的细胞相比,在3D模型中生长的细胞在增殖和衰老状态之间显示出更鲜明的对比,也影响了包括核酸在内的生物分子的丰度。两种振动光谱技术的训练精度均> 96%,这证明了这些无标记测量值适合检测3D皮肤模型中的这些细胞状态。
更新日期:2017-11-20
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