Group for Hematology and Stem Cells, Institute for Medical Research, National Institute of Republic of Serbia, University of Belgrade, 11129 Belgrade, Serbia.
Center for Solid State Physics and New Materials, Institute of Physics Belgrade, University of Belgrade, Pregrevica 118, 11080 Belgrade, Serbia.
Int J Mol Sci. 2022 Apr 28;23(9):4915. doi: 10.3390/ijms23094915.
The heterogeneity of stem cells represents the main challenge in regenerative medicine development. This issue is particularly pronounced when it comes to the use of primary mesenchymal stem/stromal cells (MSCs) due to a lack of identification markers. Considering the need for additional approaches in MSCs characterization, we applied Raman spectroscopy to investigate inter-individual differences between bone marrow MSCs (BM-MSCs). Based on standard biological tests, BM-MSCs of analyzed donors fulfill all conditions for their characterization, while no donor-related specifics were observed in terms of BM-MSCs morphology, phenotype, multilineage differentiation potential, colony-forming capacity, expression of pluripotency-associated markers or proliferative capacity. However, examination of BM-MSCs at a single-cell level by Raman spectroscopy revealed that despite similar biochemical background, fine differences in the Raman spectra of BM-MSCs of each donor can be detected. After extensive principal component analysis (PCA) of Raman spectra, our study revealed the possibility of this method to diversify BM-MSCs populations, whereby the grouping of cell populations was most prominent when cell populations were analyzed in pairs. These results indicate that Raman spectroscopy, as a label-free assay, could have a huge potential in understanding stem cell heterogeneity and sorting cell populations with a similar biochemical background that can be significant for the development of personalized therapy approaches.
干细胞的异质性是再生医学发展的主要挑战。在使用原发性间充质干细胞（MSCs）时，由于缺乏鉴定标记物，这个问题尤为突出。考虑到需要对 MSCs 进行额外的特征描述，我们应用拉曼光谱技术来研究骨髓间充质干细胞（BM-MSCs）之间的个体间差异。基于标准的生物学测试，分析供体的 BM-MSCs 符合其特征描述的所有条件，而在 BM-MSCs 形态、表型、多向分化潜能、集落形成能力、多能性相关标志物的表达或增殖能力方面没有观察到供体相关的特异性。然而，通过拉曼光谱对 BM-MSCs 进行单细胞水平的检测表明，尽管存在相似的生化背景，但仍可以检测到每个供体 BM-MSCs 的拉曼光谱存在细微差异。经过对拉曼光谱进行广泛的主成分分析（PCA）后，我们的研究表明该方法有可能使 BM-MSCs 群体多样化，当成对分析细胞群体时，细胞群体的分组最为明显。这些结果表明，拉曼光谱作为一种无标记检测方法，在理解干细胞异质性和分选具有相似生化背景的细胞群体方面具有巨大的潜力，这对于开发个性化治疗方法可能具有重要意义。
