Biomolecular phenotyping and heterogeneity assessment of mesenchymal stromal cells using label-free Raman spectroscopy

Easy, quantitative measures of biomolecular heterogeneity and high-stratified phenotyping are needed to identify and characterise complex disease processes at the single-cell level, as well as to predict cell fate. Here, we demonstrate how Raman spectroscopy can be used in the difficult-to-assess case of clonal, bone-derived mesenchymal stromal cells (MSCs) to identify MSC lines and group these according to biological function (e.g., differentiation capacity). Biomolecular stratification is achieved using high-precision measures obtained from representative statistical sampling that also enable quantified heterogeneity assessment. Application to primary MSCs and human dermal fibroblasts shows use of these measures as a label-free assay to classify cell sub-types within complex heterogeneous cell populations, thus demonstrating the potential for therapeutic translation, and broad application to the phenotypic characterisation of other cells.

Automated summary

This study demonstrates the use of Raman spectroscopy for classifying and stratifying bone-derived mesenchymal stromal cells based on biological function, with quantified assessment of biomolecular stratification and cell heterogeneity achieved through high-precision measurements and statistical sampling. The results show the potential of these methods as label-free assays for cell sub-type classification within complex heterogeneous populations, with implications for therapeutic translation and phenotypic characterisation of various cells.