Label-Free Stem Cell Differentiation Monitoring
Stem cell differentiation monitoring is crucial for directing stem cells toward specific cell lineages, essential in regenerative medicine and drug discovery. Molecular markers, imaging techniques, and functional assays help track the progression of stem cells, ensuring they transform into the desired cell types. Continuous monitoring methods are sought to overcome challenges related to heterogeneity and to provide real-time insights into the dynamics of differentiation.
Analyzing stem cell differentiation faces challenges in addressing population heterogeneity, establishing standardized criteria for biomarkers, and achieving non-invasive, real-time monitoring. Additionally, quantifying differentiated cell populations accurately remains a hurdle, particularly in complex, mixed cultures. Ongoing research focuses on innovative techniques to overcome these challenges, with the ultimate goal of enhancing the reliability and effectiveness of stem cell-based therapies and applications. While several biochemical markers exist that could be used to measure the ability of cells to self-renew, expression levels often vary and can have significant overlap between different cell lines as observed by flow cytometry. New fluorescently interrogated markers could be discovered that might aid in the specificity, but this adds additional cost and complexity to an already difficult task. Thus, a label-free method for stem cell characterization is desirable and will help drive down costs and accelerate development and personalized biomanufacturing.
Discover the Unknown Using Laser Force Cytology™
Antibodies are powerful tools, but have limitations including:
Induced phenotypic changes
False positives from non-specific binding and false negatives from non-uniform expression
Requirements for prior knowledge
Takes time and resources to develop
Unsuitable as a PAT
Real-time monitoring allows for quantitative and universal characterization of cell differentiation and the potential to discover previously unknown changes. This PAT allows for rapid knowledge throughout a lengthy production process, saving valuable time and resources, preventing costly failures, and ultimately enabling faster treatment to the patient.
Radiance® can be used to monitor iPSC and other cell differentiation pathways in a label-free manner, allowing for unbiased characterization of cells without the limitations of antibodies, enabling a rapid and label-free indication of commitment towards a specific lineage.
Quantitative Characterization of hMSC Differentiation Using Radiance®
Shown below are results illustrating LFC™ measured changes in velocity (optical force) between undifferentiated human mesenchymal stem cells (hMSCs) and their resulting differentiation into two mature cell types: osteocytes and adipocytes. When compared to undifferentiated hMSCs, the population of osteocytes shows an increase in velocity, versus a decrease in velocity in the adipocyte population. Changes are seen in multiple parameters as the cells differentiate, demonstrating the capability for Radiance® to monitor the differentiation process and providing an opportunity to use multivariate analysis to comprehensively capture and visualize the subtle changes across all the LFC™ parameters.
