MSCProfiler: a single cell image processing workflow to investigate mesenchymal stem cell heterogeneity

Single cell cytometry has demonstrated plausible immuno-heterogeneity of mesenchymal stem cells (MSCs) owing to their multivariate stromal origin. To contribute successfully to next-generation stem cell therapeutics, a deeper understanding of their cellular morphology and immunophenotype is important. In this study, the authors describe MSCProfiler, an image analysis pipeline developed using CellProfiler software. This workflow can extract geometrical and texture features such as shape, size, eccentricity and entropy, along with intensity values of the surface markers from multiple single cell images obtained using imaging flow cytometry. This screening pipeline can be used to analyze geometrical and texture features of all types of MSCs across different passages hallmarked by enhanced feature extraction potential from brightfield and fluorescent images of the cells. This study describes the development of an enhanced image feature extraction approach to analyze single cell image data of mesenchymal stem cells (MSCs). MSCProfiler, an automated image analysis pipeline, was developed using CellProfiler software to analyze geometrical and morphological/texture features and fluorescence intensities of biomarkers in MSCs across passages and tissue sources. Compared to existing image analysis tools, this workflow is devoid of human bias and is marked by the efficiency with which it filters out nontarget images and extracts features from brightfield and fluorescent images of cells.