Differential Phase Contrast Imaging to Predict MSC Immune Function

Abstract

Mesenchymal stromal cells (MSCs) exhibit significant immunomodulatory potential, making them promising candidates for cell-based therapies in autoimmune and inflammatory diseases. However, the heterogeneity of MSC cultures and a lack of robust, predictive potency assays have hindered their clinical translation. In this study, the potential of single-cell morphological imaging during MSC expansion is explored as a method to estimate indoleamine-2,3-dioxygenase (IDO) protein and enzyme activity, a common immunosuppressive capacity measure. Fluorescence and label-free quantitative differential phase contrast (qDPC) imaging is employed to non-invasively extract morphological features from live MSCs during biomanufacturing with machine learning (ML) regression models to predict single cell IDO activity. qDPC imaging characterization is extended to estimate a previously established consensus model of MSC potency based on their IDO activity and immune suppression on T cells. These findings establish a foundation for scalable, non-destructive monitoring of MSC immunomodulatory capacity, facilitating the future development of quality control strategies for MSC manufacturing and clinical applications.

Keywords: Mesenchymal stromal cells; cell morphology; immunosuppressive capacity; quantitative differentiation phase contrast imaging.