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拉曼显微光谱法:迈向对不同群体牙干细胞的更好区分与剖析

Raman microspectroscopy: toward a better distinction and profiling of different populations of dental stem cells.

作者信息

Simonović Jelena, Toljić Boško, Rašković Božidar, Jovanović Vladimir, Lazarević Miloš, Milošević Maja, Nikolić Nadja, Panajotović Radmila, Milašin Jelena

机构信息

Jelena Milašin, School of Dental Medicine, University of Belgrade, Dr Subotića 8, 11000 Belgrade, Serbia,

出版信息

Croat Med J. 2019 Apr 30;60(2):78-86. doi: 10.3325/CroatMedJ_60_0078.

Abstract

AIM

To characterize stem cells originating from different dental tissues (apical papilla [SCAP], dental follicle [DFSC], and pulp [DPSC]) and test the capacity of Raman microspectroscopy to distinguish between the three dental stem cell types.

METHODS

SCAP, DFSC, and DPSC cultures were generated from three immature wisdom teeth originating from three patients. Cell stemness was confirmed by inducing neuro-, osteo-, chondro-, and adipo-differentiaton and by mesenchymal marker expression analysis by flow-cytometry and real-time polymerase chain reaction. Cellular components were then evaluated by Raman microspectroscopy.

RESULTS

We found differences between SCAP, DFSC, and DPSC Raman spectra. The ratio between proteins and nucleic acids (748/770), a parameter for discriminating more differentiated from less differentiated cells, showed significant differences between the three cell types. All cells also displayed a fingerprint region in the 600-700 cm-1 range, and characteristic lipid peaks at positions 1440 cm-1 and 1650 cm-1.

CONCLUSION

Although different dental stem cells exhibited similar Raman spectra, the method enabled us to make subtle distinction between them.

摘要

目的

鉴定源自不同牙组织(根尖乳头干细胞[SCAP]、牙囊干细胞[DFSC]和牙髓干细胞[DPSC])的干细胞,并测试拉曼光谱法区分这三种牙干细胞类型的能力。

方法

从三名患者的三颗未成熟智齿中培养出SCAP、DFSC和DPSC。通过诱导神经、骨、软骨和脂肪分化以及通过流式细胞术和实时聚合酶链反应进行间充质标志物表达分析来确认细胞干性。然后通过拉曼光谱法评估细胞成分。

结果

我们发现SCAP、DFSC和DPSC的拉曼光谱存在差异。蛋白质与核酸的比率(748/770)是区分分化程度较高与较低细胞的一个参数,在这三种细胞类型之间显示出显著差异。所有细胞在600 - 700 cm-1范围内也显示出一个指纹区域,以及在1440 cm-1和1650 cm-1位置的特征性脂质峰。

结论

尽管不同的牙干细胞表现出相似的拉曼光谱,但该方法使我们能够对它们进行细微区分。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6f26/6509629/a4ee95c3c934/CroatMedJ_60_0078-F1.jpg

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