Quantitative Phase Microscopy for the evaluation and prediction of Mesenchymal Stromal Cell osteogenesis

FDA-approved mesenchymal stromal cell (MSC) clinical therapies have been elusive despite strong evidence of their applications in regenerative medicine due to their potential for multi-lineage differentiation, immunomodulation, and paracrine factor secretion. MSCs can provide regenerative potential to areas of the body such as tendons, ligaments, cartilage, and bone. A major hurdle in translation is determining cell quality during expansion and biomanufacturing, so methods that can robustly and non-destructively evaluate their performance will significantly advance this field. With the development of advanced imaging-based techniques to access high-dimensional morphological analysis, we investigated short-term morphological profiles of MSCs correlated with long-term osteogenic differentiation. Using label-free phase imaging, we captured features like GLCM contrast and correlation that tracked differentiation over time. UMAP confirmed visual trends, and random forest models successfully predicted day 28 mineralization from early features. These findings lay the groundwork for future in-line monitoring and selection of high-performance MSC cultures during manufacturing.