Abstract
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.
Highlights
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
The mesenchymal stromal cells (MSCs) lines in this study were generated by isolating single-cell-derived colonies from a heterogeneous MSC population that was immortalised with human telomerase reverse transcriptase[14]
The Raman spectra for the human telomerase reverse transcriptase (hTERT) MSCs were tested for celltype discrimination and the ability to group Y101/Y201 vs. Y102/Y202 due to these groups having previously defined, shared biological features, such as morphologies, immunomodulatory capacities and differentiation competencies[14]
Summary
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. I.e., phenotypic plasticity, may result from treatment-related genetic-scale c hanges[11] From these examples alone, the enormity in developing tractable, quantitative assessment and phenotyping of cell-level and population-level heterogeneity, and the relationship of these factors to accurate assay development, is apparent. The enormity in developing tractable, quantitative assessment and phenotyping of cell-level and population-level heterogeneity, and the relationship of these factors to accurate assay development, is apparent To assist with these challenges, spectroscopic methods, such as Raman spectroscopy (RS), are being developed and assessed as complementary routes for molecular-level characterisation of intra- and inter-cellular properties. Its use in translational biomedical applications could be further expanded by the development of easy-to-prepare Ramanbased assays for stratified phenotyping, created from rigorous data analyses, and verified by reproducibility tests via robust statistical assessment
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