Abstract
Techniques for following the differentiation of human mesenchymal stem cells (hMSCs) in laboratory settings prior to their transplantation into living organisms are essential for progress in tissue engineering and regenerative medicine. In this study, we have used a non-toxic fluorescent dye (ChromaLIVE™) coupled with an artificial intelligence (AutoHCS™) powered image analysis system for real-time monitoring of the differentiation of hMSCs. To validate the performance of this novel Live-Cell Imaging assay, its accuracy was benchmarked to a well-established immunocytochemistry method for studying MSC differentiation into osteoblasts. This innovative method utilizes the distinctive phenotypic signature detected by the non-toxic dye to identify and measure differentiation phenotypes, which were found to align with the expression of osteogenic markers. As a highly sensitive, affordable, non-destructive and scalable kinetic assay, this new technology offers promise as a dependable tool for monitoring stem cell differentiation. By delivering real-time insights into the quality of cell batches, it facilitates prompt adjustments and optimization of culture conditions.